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> Mouse TrueBlot® ULTRA: Anti-Mouse Ig HRP

Mouse TrueBlot® ULTRA: Anti-Mouse Ig HRP

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Cat# 18-8817-33

Size : 200ul

Brand : Rockland Immunochemicals

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Phone : +1 850 650 7790

Specifications for Mouse TrueBlot® ULTRA: Anti-Mouse Ig HRP

Product Details

Mouse TrueBlot® ULTRA: Anti-Mouse Ig HRP - 18-8817-30
Anti-Mouse IgG HRP, TrueBlot, HRP TrueBlot ULTRA, Peroxidase TrueBlot, TrueBlot for IP/WB, TrueBlot for immunoprecipitation, TrueBlot for western blotting
Rat
Peroxidase (HRP) ULTRA
Monoclonal
IgG

Target Details

Mouse
Mouse TrueBlot® ULTRA Antibody Peroxidase Conjugate was prepared from tissue culture supernatant by Protein G affinity chromatography. Assay by immunoelectrophoresis resulted in a single precipitin arc against Anti-Mouse Serum. Reactivity is observed against native Mouse IgG by both Western blot and ELISA.

Application Details

ELISA, IP, WB
Biochemical Assay, ChIP, IF  - View References
Mouse TrueBlot® ULTRA has been tested in ELISA, IP, and Western blot and may also be used for detection in immunoblotting assays that do not employ immunoprecipitation. Mouse IgG TrueBlot® ULTRA is provided as 1000X solution. To achieve best results when detecting mouse IgG1 subtypes, we recommend performing a dot blot or titration to determine the optimal dilution factor for your desired application. All recommended dilutions for listed applications are intended as an initial recommendation, specific conditions for each protein and antibody combination should be specifically optimized by the end user. In order to conserve reagent, we recommend incubating the blots from minigels in sealed bags (removing as much air as possible) with minimal volume (2-3 mLs). If used conservatively at 2.5mls/blot will yield enough reagent for 8 blots. Note that there are three key procedural considerations: 1. Protein A or G beads may be used with the mouse, goat and sheep TrueBlot secondaries, but not with the rabbit TrueBlot secondary. Use of protein A or G beads with the rabbit TrueBlot will result in contaminating bands. 2. Immunoprecipitate should be completely reduced. 3. BLOTTO/Milk should be used as the blocking protein for the immunoblot.

Formulation

1.0 mg/mL
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Proclin is added as an antimicrobial agent.
0.1 mg/ml Bovine Serum Albumin (BSA) - IgG and Protease free, 50% (v/v) Glycerol

Shipping & Handling

Wet Ice
Store at -20°C. This product is guaranteed for 6 months upon receipt, when handled and stored as instructed.
Expiration date is six (6) months from date of receipt.

Background

Mouse IgG TrueBlot® ULTRA is a unique Anti-Mouse IgG monoclonal secondary antibody. Mouse IgG TrueBlot® ULTRA enables detection of immunoblotted target protein bands, without hindrance by interfering immunoprecipitating immunoglobulin heavy and light chains. It is easy to generate publication-quality IP/Western blot data with Mouse IgG TrueBlot® ULTRA, simply substitute the conventional Anti-Mouse IgG blotting reagent with Mouse IgG TrueBlot® ULTRA and follow the prescribed protocol for sample preparation and immunoblotting. Mouse IgG TrueBlot® ULTRA is ideal for use in protocols involving immunoblotting of immunoprecipitated proteins. Mouse IgG TrueBlot® ULTRA preferentially detects the non-reduced form of mouse IgG (IgG1, IgG2a, IgG2b, IgG3) over the reduced, SDS-denatured form of IgG. When the immunoprecipitate is fully reduced immediately prior to SDS-gel electrophoresis, reactivity of Mouse IgG TrueBlot® ULTRA with the 55 kDa heavy chains and the 23 kDa light chains of the immunoprecipitating antibody is minimized thereby eliminating interference by the heavy and light chains of the immunoprecipitating antibody in IP/immunoblot applications. Applications include studies examining post-translational modification (e.g., phosphorylation or acetylation) or protein-protein interactions.

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PubMed
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WB, IB, PCA
PubMed
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IP, Co-IP
PubMed
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(). Identification of a Small-Molecule Inhibitor That Disrupts the SIX1/EYA2 Complex, EMT, and Metastasis. Cancer Res.
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PubMed
(). KAP1 Is a Chromatin Reader that Couples Steps of RNA Polymerase II Transcription to Sustain Oncogenic Programs. Mol Cell.
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IP, Co-IP
PubMed
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WB, IB, PCA
PubMed
(). Protective and inhibitory effects of acteoside from Abeliophyllum distichum Nakai against oxidative DNA damage. Mol Med Rep.
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WB, IB, PCA
PubMed
(). eIF2B as a Target for Viral Evasion of PKR-Mediated Translation Inhibition. mBio.
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WB, IB, PCA
PubMed
(). Targeting Opsin4/Melanopsin with a Novel Small Molecule Suppresses PKC/RAF/MEK/ERK Signaling and Inhibits Lung Adenocarcinoma Progression. Mol Cancer Res.
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WB, IB, PCA
PubMed
(). Pooled CRISPR screens with imaging on microraft arrays reveals stress granule-regulatory factors. Nat Methods.
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WB, IB, PCA
PubMed
(). A Comprehensive Map of the Monocyte-Derived Dendritic Cell Transcriptional Network Engaged upon Innate Sensing of HIV. Cell Rep.
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WB, IB, PCA
PubMed
(). An inventory of interactors of the human HSP60/HSP10 chaperonin in the mitochondrial matrix space. Cell Stress Chaperones.
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WB, IB, PCA
PubMed
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WB, IB, PCA
PubMed
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PubMed
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WB, IB, PCA
PubMed
(). Periostin and tenascin-C interaction promotes angiogenesis in ischemic proliferative retinopathy. Sci Rep.
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IP, Co-IP; WB, IB, PCA
PubMed
(). Fbxo7 and Pink1 play a reciprocal role in regulating their protein levels. Aging (Albany NY).
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WB, IB, PCA
PubMed
(). Oxidative Stress Culminates in Intracellular Bacteria Expulsion Via NRF2/KEAP1 Induction of RAB27B. Cell Press Preprint
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WB, IB, PCA
Source
(). WSTF acetylation by MOF promotes WSTF activities and oncogenic functions. Oncogene.
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PubMed
(). Gene-and Species-Specific hox mRNA translation by ribosome expansion segments. Mol Cell.
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WB, IB, PCA
PubMed
(). TREX2 Exonuclease Causes Spontaneous Mutations and Stress-Induced Replication Fork Defects in Cells Expressing RAD51K133A. Cell Rep.
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WB, IB, PCA
PubMed
(). Tristetraprolin promotes hepatic inflammation and tumor initiation but restrains cancer progression to malignancy. Cell Mol Gastroenterol Hepatol.
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WB, IB, PCA
PubMed
(). Inhibiting calpain 1 and 2 in cyclin G associated kinase–knockout mice mitigates podocyte injury. JCI Insight.
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WB, IB, PCA
PubMed
(). Bone-Specific Overexpression of PITX1 Induces Senile Osteoporosis in Mice Through Deficient Self-Renewal of Mesenchymal Progenitors and Wnt Pathway Inhibition. Sci Rep.
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WB, IB, PCA
PubMed
(). The Crohn's Disease Risk Factor IRGM Limits NLRP3 Inflammasome Activation by Impeding Its Assembly and by Mediating Its Selective Autophagy. Molecular Cell
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WB, IB, PCA
PubMed
(). The Crohn's Disease Risk Factor IRGM Limits NLRP3 Inflammasome Activation by Impeding Its Assembly and by Mediating Its Selective Autophagy. Molecular Cell
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IP, Co-IP
PubMed
(). Broad-spectrum receptor tyrosine kinase inhibitors overcome de novo and acquired modes of resistance to EGFR-targeted therapies in colorectal cancer. Oncotarget
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WB, IB, PCA
PubMed
(). Microbial recognition by GEF-H1 controls IKKε mediated activation of IRF5. Nature Communications
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IP, Co-IP
PubMed
(). Apurinic/apyrimidinic endonuclease 1 (APE1) is dispensable for activation-induced cytidine deaminase (AID)-dependent somatic hypermutation in the immunoglobulin gene. Int Immunol.
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WB, IB, PCA
PubMed
(). The Hippo Pathway Blocks Mammalian Retinal Müller Glial Cell Reprogramming. Cell Rep.
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IP, Co-IP
PubMed
(). BIN1 favors the spreading of Tau via extracellular vesicles. Sci Rep.
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WB, IB, PCA
PubMed
(). BIN1 favors the spreading of Tau via extracellular vesicles. Sci Rep.
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WB, IB, PCA; IP, Co-IP
PubMed
(). Unique and Shared Epigenetic Programs of the CREBBP and EP300 Acetyltransferases in Germinal Center B Cells Reveal Targetable Dependencies in Lymphoma. Immunity.
Applications
WB, IB, PCA
PubMed
(). Parkin truncating variants result in a loss-of-function phenotype. Sci Rep.
Applications
WB, IB, PCA; IP, Co-IP
PubMed
(). YAP/TAZ Activation Drives Uveal Melanoma Initiation and Progression. Cell Rep.
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WB, IB, PCA
PubMed
(). The Transcription Factor MAZR/PATZ1 Regulates the Development of FOXP3+ Regulatory T Cells. Cell Rep.
Applications
WB, IB, PCA
PubMed
(). A trimeric Rab7 GEF controls NPC1-dependent lysosomal cholesterol export. Nat Commun
Applications
IF, Confocal Microscopy
PubMed
(). p53-Pirh2 Complex Promotes Twist1 Degradation and Inhibits EMT. Mol Cancer Res.
Applications
WB, IB, PCA
PubMed
(). Protein kinase Cα-mediated phosphorylation of Twist1 at Ser-144 prevents Twist1 ubiquitination and stabilizes it. J Biol Chem.
Applications
WB, IB, PCA
PubMed
(). TAM Kinases Promote Necroptosis by Regulating Oligomerization of MLKL. Mol Cell.
Applications
WB, IB, PCA
PubMed
(). Irradiated Human Fibroblasts as a Substitute Feeder Layer to Irradiated Mouse 3T3 for the Culture of Human Corneal Epithelial Cells: Impact on the Stability of the Transcription Factors Sp1 and NFI. Int J Mol Sci.
Applications
WB, IB, PCA
PubMed
(). Pink1 regulates FKBP5 interaction with AKT/PHLPP and protects neurons from neurotoxin stress induced by MPP. J Neurochem.
Applications
WB, IB, PCA
PubMed
(). HIV-1 Envelope Overcomes NLRP3-Mediated Inhibition of F-Actin Polymerization for Viral Entry. Cell Rep.
Applications
WB, IB, PCA
PubMed
(). Epigenetic modulators hydralazine and sodium valproate act synergistically in VEGI-mediated anti-angiogenesis and VEGF interference in human osteosarcoma and vascular endothelial cells. Int J Oncol.
Applications
WB, IB, PCA
PubMed
(). Acquired Resistance of EGFR-Mutated Lung Cancer to Tyrosine Kinase Inhibitor Treatment Promotes PARP Inhibitor Sensitivity. Cell Rep.
Applications
WB, IB, PCA
PubMed
(). The Myb-MuvB Complex Is Required for YAP-Dependent Transcription of Mitotic Genes. Cell Rep.
Applications
WB, IB, PCA
PubMed
(). Expression of protein disulfide isomerase A3 and its clinicopathological association in gastric cancer. Oncol Rep.
Applications
WB, IB, PCA
PubMed
(). Dual Sensing of Physiologic pH and Calcium by EFCAB9 Regulates Sperm Motility. Cell.
Applications
WB, IB, PCA
PubMed
(). Cand1-Mediated Adaptive Exchange Mechanism Enables Variation in F-Box Protein Expression. Molecular Cell
Applications
WB, IB, PCA
PubMed
(). Effect of human very low-density lipoproteins on cardiotrophin-like cytokine factor 1 (CLCF1) activity. Scientific Reports
Applications
IP, Co-IP
PubMed
(). The miR-106b-25 cluster mediates breast tumor initiation through activation of NOTCH1 via direct repression of NEDD4L. Oncogene
Applications
IP, Co-IP
PubMed
(). ATP depletion during mitotic arrest induces mitotic slippage and APC/CCdh1-dependent cyclin B1 degradation. Experimental & Molecular Medicine
Applications
IP, Co-IP; WB, IB, PCA
PubMed
(). IL1RAP potentiates multiple oncogenic signaling pathways in AML. Journal of Experimental Medicine
Applications
IP, Co-IP
PubMed
(). Lipid-dependent deposition of alpha-synuclein and Tau on neuronal Secretogranin II-positive vesicular membranes with age. Scientific Reports
Applications
WB, IB, PCA
PubMed
(). PDI Knockdown Inhibits Seizure Activity in Acute Seizure and Chronic Epilepsy Rat Models via S-Nitrosylation-Independent Thiolation on NMDA Receptor. Frontiers in Cellular Neuroscience
Applications
WB, IB, PCA
PubMed
(). The sterol-responsive RNF145 E3 ubiquitin ligase mediates the degradation of HMG-CoA reductase together with gp78 and Hrd1. Elife
Applications
IP, Co-IP
PubMed
(). Gankyrin Promotes Tumor-Suppressor Protein Degradation to Drive Hepatocyte Proliferation. Cellular and Molecular Gastroenterology and Hepatology
Applications
WB, IB, PCA
PubMed
(). Phase transitioned nuclear Oskar promotes cell division of Drosophila primordial germ cells. Elife
Applications
WB, IB, PCA
PubMed
(). Extracellular Protein Fibulin-7 and Its C-Terminal Fragment Have In Vivo Antiangiogenic Activity. Scientific Reports
Applications
WB, IB, PCA
PubMed
(). Sestrin2 facilitates glutamine-dependent transcription of PGC-1α and survival of liver cancer cells under glucose limitation. FEBS J.
Applications
WB, IB, PCA; IP, Co-IP
PubMed
(). lncRNA Epigenetic Landscape Analysis Identifies EPIC1 as an Oncogenic lncRNA that Interacts with MYC and Promotes Cell-Cycle Progression in Cancer. Cancer Cell.
Applications
WB, IB, PCA
PubMed
(). Modulation of Protein-Interaction States through the Cell Cycle. Cell.
Applications
IP, Co-IP
PubMed
(). PELI1 Selectively Targets Kinase-Active RIP3 for Ubiquitylation-Dependent Proteasomal Degradation. Mol Cell.
Applications
WB, IB, PCA; IP, Co-IP
PubMed
(). 53BP1 cooperation with the REV7-shieldin complex underpins DNA structure-specific NHEJ. Nature.
Applications
WB, IB, PCA
PubMed
(). MEF2B Instructs Germinal Center Development and Acts as an Oncogene in B Cell Lymphomagenesis. Cancer Cell.
Applications
WB, IB, PCA
PubMed
(). The interaction between IKKα and LC3 promotes type I interferon production through the TLR9-containing LAPosome. Sci Signal.
Applications
WB, IB, PCA
PubMed
(). Active N6-Methyladenine Demethylation by DMAD Regulates Gene Expression by Coordinating with Polycomb Protein in Neurons. Mol Cell.
Applications
WB, IB, PCA
PubMed
(). Flap endonuclease 1 is involved in cccDNA formation in the hepatitis B virus. PLoS Pathog.
Applications
WB, IB, PCA
PubMed
(). Expression of Human Cytomegalovirus IE1 Leads to Accumulation of Mono-SUMOylated PML That Is Protected from Degradation by Herpes Simplex Virus 1 ICP0. J Virol.
Applications
WB, IB, PCA
PubMed
(). NONO Detects the Nuclear HIV Capsid to Promote cGAS-Mediated Innate Immune Activation. Cell. 2018;175(2):488-501.e22. doi:10.1016/j.cell.2018.08.062 Cell.
Applications
WB, IB, PCA
PubMed
(). The RNA-binding protein PTBP1 is necessary for B cell selection in germinal centers. Nat Immunol.
Applications
WB, IB, PCA
PubMed
(). Original Ligand for LTβR Is LIGHT: Insight into Evolution of the LT/LTβR System. J Immunol.
Applications
WB, IB, PCA; In Vitro Binding Assay
PubMed
(). The multi-subunit GID/CTLH E3 ubiquitin ligase promotes cell proliferation and targets the transcription factor Hbp1 for degradation. Elife.
Applications
WB, IB, PCA
PubMed
(). Identification of hepatic protein-protein interaction targets for betaine homocysteine S-methyltransferase. PLoS One.
Applications
WB, IB, PCA
PubMed
(). Inhibition of TRAF6 ubiquitin-ligase activity by PRDX1 leads to inhibition of NFKB activation and autophagy activation. Autophagy.
Applications
WB, IB, PCA
PubMed
(). Elucidating the mitochondrial proteome of Toxoplasma gondii reveals the presence of a divergent cytochrome c oxidase. Elife.
Applications
WB, IB, PCA
PubMed
(). Cytoplasmic E2f4 forms organizing centres for initiation of centriole amplification during multiciliogenesis. Nat Commun.
Applications
WB, IB, PCA
PubMed
(). Protein Phosphotyrosine Phosphatase 1B (PTP1B) in Calpain-dependent Feedback Regulation of Vascular Endothelial Growth Factor Receptor (VEGFR2) in Endothelial Cells: IMPLICATIONS IN VEGF-DEPENDENT ANGIOGENESIS AND DIABETIC WOUND HEALING. Journal of Biological Chemistry
Applications
IP, Co-IP
PubMed
(). A Cytoplasmic RNA Virus Alters the Function of the Cell Splicing Protein SRSF2. Journal of Virology
Applications
IP, Co-IP
PubMed
(). Interference of the complex between NCS-1 and Ric8a with phenothiazines regulates synaptic function and is an approach for fragile X syndrome. Proc. Natl. Acad. Sci. U.S.A
Applications
IP, Co-IP; WB, IB, PCA
PubMed
(). An interaction domain in human SAMD9 is essential for myxoma virus host-range determinant M062 antagonism of host anti-viral function. Virology
Applications
IP, Co-IP; WB, IB, PCA
PubMed
(). NLRP1 promotes tumor growth by enhancing inflammasome activation and suppressing apoptosis in metastatic melanoma. Oncogene
Applications
IP, Co-IP; WB, IB, PCA
PubMed
(). Human mitochondrial cytochrome c oxidase assembly factor COX18 acts transiently as a membrane insertase within the subunit 2 maturation module. Journal of Biological Chemistry
Applications
IP, Co-IP; WB, IB, PCA
PubMed
(). An FAK-YAP-mTOR Signaling Axis Regulates Stem Cell-Based Tissue Renewal in Mice. Cell Stem Cell
Applications
IF, Confocal Microscopy; WB, IB, PCA
PubMed
(). RNA Binding Protein CUGBP1 Inhibits Liver Cancer in a Phosphorylation-Dependent Manner. Molecular and Cellular Biology
Applications
WB, IB, PCA
PubMed
(). BAP1 regulates IP3R3-mediated Ca2+ flux to mitochondria suppressing cell transformation. Nature
Applications
WB, IB, PCA
PubMed
(). Cytoplasmic E2f4 forms organizing centres for initiation of centriole amplification during multiciliogenesis. Nature Communications
Applications
IP, Co-IP
PubMed
(). Centriolar Satellites Control GABARAP Ubiquitination and GABARAP-Mediated Autophagy. Current Biology
Applications
IP, Co-IP
PubMed
(). A Brucella Type IV Effector Targets the COG Tethering Complex to Remodel Host Secretory Traffic and Promote Intracellular Replication. Cell Host Microbe
Applications
WB, IB, PCA
PubMed
(). A Brucella Type IV Effector Targets the COG Tethering Complex to Remodel Host Secretory Traffic and Promote Intracellular Replication. Cell Host Microbe
Applications
IF, Confocal Microscopy
PubMed
(). POPX2 phosphatase regulates apoptosis through the TAK1-IKK-NF-κB pathway. Cell Death & Disease
Applications
WB, IB, PCA
PubMed
(). Blue light and CO2 signals converge to regulate light-induced stomatal opening. Nature Communications
Applications
IP, Co-IP; WB, IB, PCA
PubMed
(). Mechanistic Insights into Autoinhibition of the Oncogenic Chromatin Remodeler ALC1. Molecular Cell
Applications
IP, Co-IP
PubMed
(). Cell cycle-dependent phosphorylation regulates RECQL4 pathway choice and ubiquitination in DNA double-strand break repair. Nature Communications
Applications
IP, Co-IP; WB, IB, PCA
PubMed
(). Zinc finger protein ZPR9 functions as an activator of AMPK-related serine/threonine kinase MPK38/MELK involved in ASK1/TGF-β/p53 signaling pathways. Scientific Reports
Applications
IP, Co-IP; WB, IB, PCA
PubMed
(). Angiotensin II activates CaV1.2 Ca2+channels through β-arrestin2 and casein kinase 2 in mouse immature cardiomyocytes. The Journal of Physiology
Applications
WB, IB, PCA
PubMed
(). PrimPol is required for replication reinitiation after mtDNA damage. Proc Natl Acad Sci U S A.
Applications
WB, IB, PCA
PubMed
(). Composition and Regulation of the Cellular Repertoire of SCF Ubiquitin Ligases. Cell.
Applications
WB, IB, PCA
PubMed
(). SFPQ associates to LSD1 and regulates the migration of newborn pyramidal neurons in the developing cerebral cortex. Int J Dev Neurosci.
Applications
WB, IB, PCA
PubMed
(). The Mammalian Ribo-interactome Reveals Ribosome Functional Diversity and Heterogeneity. Cell.
Applications
WB, IB, PCA
PubMed
(). Human cytomegalovirus-encoded viral cyclin-dependent kinase (v-CDK) UL97 phosphorylates and inactivates the retinoblastoma protein-related p107 and p130 proteins. J Biol Chem.
Applications
WB, IB, PCA
PubMed
(). Phosphorylation of transcriptional regulators in the retinoblastoma protein pathway by UL97, the viral cyclin-dependent kinase encoded by human cytomegalovirus. Virology.
Applications
WB, IB, PCA
PubMed
(). A Short cis-Acting Motif in the M112-113 Promoter Region Is Essential for IE3 To Activate M112-113 Gene Expression and Is Important for Murine Cytomegalovirus Replication. J Virol.
Applications
WB, IB, PCA
PubMed
(). Contribution of JAK2 mutations to T-cell lymphoblastic lymphoma development. Leukemia
Applications
IP, Co-IP
PubMed
(). The Cellular Protein Complex Associated with a Transforming Region of E1A Contains c-MYC. Journal of Virology
Applications
WB, IB, PCA
PubMed
(). The Cellular Protein Complex Associated with a Transforming Region of E1A Contains c-MYC. Journal of Virology
Applications
WB, IB, PCA; IP, Co-IP
PubMed
(). Characterization of RyDEN (C19orf66) as an Interferon-Stimulated Cellular Inhibitor against Dengue Virus Replication. PLOS Pathogens
Applications
WB, IB, PCA
PubMed
(). Leucine zipper motif in RRS1 is crucial for the regulation of Arabidopsis dual resistance protein complex RPS4/RRS1. Scientific Reports
Applications
IP, Co-IP; WB, IB, PCA
PubMed
(). NBR1 enables autophagy-dependent focal adhesion turnover. Journal of Cell Biology
Applications
IP, Co-IP
PubMed
(). Oral administration of Aloe vera gel powder prevents UVB-induced decrease in skin elasticity via suppression of overexpression of MMPs in hairless mice. Bioscience, Biotechnology, and Biochemistry
Applications
WB, IB, PCA
PubMed
(). Direct Demonstration That Loop1 of Scap Binds to Loop7: A CRUCIAL EVENT IN CHOLESTEROL HOMEOSTASIS. Journal of Biological Chemistry
Applications
WB, IB, PCA
PubMed
(). Cyclic Nucleotide Control of Microtubule Dynamics for Axon Guidance. The Journal of Neuroscience
Applications
WB, IB, PCA
PubMed
(). Complement system activation contributes to the ependymal damage induced by microbial neuraminidase. Journal of Neuroinflammation
Applications
WB, IB, PCA
PubMed
(). RNA Exosome Complex Regulates Stability of the Hepatitis B Virus X-mRNA Transcript in a Non-stop-mediated (NSD) RNA Quality Control Mechanism. Journal of Biological Chemistry
Applications
WB, IB, PCA
PubMed
(). SMG7 is a critical regulator of p53 stability and function in DNA damage stress response. Cell Discovery
Applications
WB, IB, PCA
PubMed
(). Cereblon negatively regulates TLR4 signaling through the attenuation of ubiquitination of TRAF6. Cell Death & Disease
Applications
WB, IB, PCA
PubMed
(). Suppressor of Cytokine Signaling (SOCS)1 Regulates Interleukin-4 (IL-4)-activated Insulin Receptor Substrate (IRS)-2 Tyrosine Phosphorylation in Monocytes and Macrophages via the Proteasome. Journal of Biological Chemistry
Applications
IP, Co-IP
PubMed
(). Eph-mediated tyrosine phosphorylation of citron kinase controls abscission. Journal of Cell Biology
Applications
IP, Co-IP; WB, IB, PCA
PubMed
(). Rac1-mediated cytoskeleton rearrangements induced by intersectin-1s deficiency promotes lung cancer cell proliferation, migration and metastasis. Molecular Cancer
Applications
IP, Co-IP; WB, IB, PCA
PubMed
(). Diverse mechanisms of metaeffector activity in an intracellular bacterial pathogen, Legionella pneumophila. Molecular Systems Biology
Applications
WB, IB, PCA
PubMed
(). Activation of the thrombopoietin receptor by mutant calreticulin in CALR-mutant myeloproliferative neoplasms. Blood.
Applications
WB, IB, PCA
PubMed
(). TRIP13 promotes error-prone nonhomologous end joining and induces chemoresistance in head and neck cancer. Nat Commun.
Applications
WB, IB, PCA
PubMed
(). Loss of the antioxidant enzyme CuZnSOD (Sod1) mimics an age-related increase in absolute mitochondrial DNA copy number in the skeletal muscle. Age (Dordr).
Applications
WB, IB, PCA
PubMed
(). The Oncogene PDRG1 Is an Interaction Target of Methionine Adenosyltransferases. PLoS One.
Applications
WB, IB, PCA
PubMed
(). A multi-stage process including transient polyploidization and EMT precedes the emergence of chemoresistent ovarian carcinoma cells with a dedifferentiated and pro-inflammatory secretory phenotype. Oncotarget
Applications
WB, IB, PCA
PubMed
(). Myotubularin-related proteins 3 and 4 interact with polo-like kinase 1 and centrosomal protein of 55 kDa to ensure proper abscission. Molecular & Cellular Proteomics
Applications
IP, Co-IP
PubMed
(). A monoclonal antibody that tracks endospore formation in the microsporidium Nosema bombycis. PLOS One
Applications
WB, IB, PCA
PubMed
(). Ebola Virus Glycoprotein Promotes Enhanced Viral Egress by Preventing Ebola VP40 From Associating With the Host Restriction Factor BST2/Tetherin. The Journal of Infectious Diseases
Applications
IP, Co-IP; WB, IB, PCA
PubMed
(). Lipopolysaccharide Induces Alveolar Macrophage Necrosis via CD14 and the P2X7 Receptor Leading to Interleukin-1α Release. Immunity
Applications
IP, Co-IP
PubMed
(). TRAF1 Coordinates Polyubiquitin Signaling to Enhance Epstein-Barr Virus LMP1-Mediated Growth and Survival Pathway Activation. PLOS Pathogens
Applications
WB, IB, PCA
PubMed
(). Hypertension-causing Mutations in Cullin3 Protein Impair RhoA Protein Ubiquitination and Augment the Association with Substrate Adaptors. Journal of Biological Chemistry
Applications
IP, Co-IP; WB, IB, PCA
PubMed
(). Differential Effects of Hepatocyte Nuclear Factor 4α Isoforms on Tumor Growth and T-Cell Factor 4/AP-1 Interactions in Human Colorectal Cancer Cells. Molecular and Cellular Biology
Applications
ChIP; WB, IB, PCA
PubMed
(). Loss of p53 enhances the function of the endoplasmic reticulum through activation of the IRE1α/XBP1 pathway. Oncotarget
Applications
WB, IB, PCA
PubMed
(). MAZR and Runx Factors Synergistically Repress ThPOK during CD8+ T Cell Lineage Development. Journal of Immunology
Applications
IP, Co-IP; WB, IB, PCA
PubMed
(). MAZR and Runx Factors Synergistically Repress ThPOK during CD8+ T Cell Lineage Development. Journal of Immunology
Applications
IP, Co-IP
PubMed
(). Expression of human carcinoembryonic antigen-related cell adhesion molecule 6 and alveolar progenitor cells in normal and injured lungs of transgenic mice. Physiological Reports
Applications
WB, IB, PCA
PubMed
(). TGF-β suppression of HBV RNA through AID-dependent recruitment of an RNA exosome complex. PLoS Pathog.
Applications
WB, IB, PCA
PubMed
(). Silencing of PMEPA1 accelerates the growth of prostate cancer cells through AR, NEDD4 and PTEN. Oncotarget.
Applications
WB, IB, PCA
PubMed
(). Molecular Determinants for the Inactivation of the Retinoblastoma Tumor Suppressor by the Viral Cyclin-dependent Kinase UL97. J Biol Chem.
Applications
WB, IB, PCA
PubMed
(). Role of extracellular matrix in development of skeletal muscle and postmortem aging of meat. Meat Sci.
Applications
WB, IB, PCA
PubMed
(). The conserved Trp114 residue of thioredoxin reductase 1 has a redox sensor-like function triggering oligomerization and crosslinking upon oxidative stress related to cell death. Cell Death Dis.
Applications
WB, IB, PCA
PubMed
(). Differential regulation of S-region hypermutation and class switch recombination by noncanonical functions of uracil DNA glycosylase. Proc Natl Acad Sci USA
Applications
ChIP; IP, Co-IP
PubMed
(). The nuclear form of glutathione peroxidase 4 colocalizes and directly interacts with protamines in the nuclear matrix during mouse sperm chromatin assembly. Spermatogenesis.
Applications
WB, IB, PCA
PubMed
(). OFD1 and flotillins are integral components of a ciliary signaling protein complex organized by polycystins in renal epithelia and odontoblasts. PLoS One.
Applications
WB, IB, PCA
PubMed
(). Specificity of anti-tau antibodies when analyzing mice models of Alzheimer's disease: problems and solutions. PLoS One.
Applications
WB, IB, PCA
PubMed
(). The COP II adaptor protein TMED7 is required to initiate and mediate the delivery of TLR4 to the plasma membrane. Sci Signal.
Applications
WB, IB, PCA
PubMed
(). Lipoprotein lipase (LPL) is associated with neurite pathology and its levels are markedly reduced in the dentate gyrus of Alzheimer's disease brains. The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
Applications
WB, IB, PCA
PubMed
(). RAB8B is required for activity and caveolar endocytosis of LRP6. Cell Rep.
Applications
WB, IB, PCA
PubMed
(). Sfrp5 modulates both Wnt and BMP signaling and regulates gastrointestinal organogenesis [corrected] in the zebrafish, Danio rerio. PLoS One.
Applications
WB, IB, PCA
PubMed
(). Aerobic exercise + weight loss decreases skeletal muscle myostatin expression and improves insulin sensitivity in older adults. Obesity (Silver Spring).
Applications
WB, IB, PCA
PubMed
(). SARM1 and TRAF6 bind to and stabilize PINK1 on depolarized mitochondria. Mol Biol Cell.
Applications
WB, IB, PCA
PubMed
(). High-content analysis of proapoptotic EphA4 dependence receptor functions using small-molecule libraries. J Biolmol Screen
Applications
WB, IB, PCA
PubMed
(). MPS1/Mph1 phosphorylates the kinetochore protein KNL1/Spc7 to recruit SAC components. Nat Cel Biol
Applications
WB, IB, PCA
PubMed
(). Host genetics and Chlamydia disease: prediction and validation of disease severity mechanisms. PLoS One.
Applications
IP, Co-IP
PubMed
(). Leucine zipper domain is required for Kaposi sarcoma-associated herpesvirus (KSHV) K-bZIP protein to interact with histone deacetylase and is important for KSHV replication. J Biol Chem.
Applications
WB, IB, PCA
PubMed
(). Growth differentiation factor-15 deficiency inhibits atherosclerosis progression by regulating interleukin-6-dependent inflammatory response to vascular injury. J Am Heart Assoc.
Applications
WB, IB, PCA
PubMed
(). H2B homology region of major immediate-early protein 1 is essential for murine cytomegalovirus to disrupt nuclear domain 10, but is not important for viral replication in cell culture. J Gen Virol.
Applications
WB, IB, PCA
PubMed
(). c-Jun N-terminal kinase phosphorylates DCP1a to control formation of P bodies. J Cell Biol.
Applications
WB, IB, PCA
PubMed
(). Ins(1,4,5)P3 receptor-mediated Ca2+ signaling and autophagy induction are interrelated. Autophagy.
Applications
WB, IB, PCA
PubMed
(). A genome-wide profiling of the humoral immune response to Chlamydia trachomatis infection reveals vaccine candidate antigens expressed in humans. J Immunol.
Applications
WB, IB, PCA
PubMed
(). Posttranslational regulation of membrane type 1-matrix metalloproteinase (MT1-MMP) in mouse PTEN null prostate cancer cells: Enhanced surface expression and differential O-glycosylation of MT1-MMP. Biochim Biophys Acta.
Applications
WB, IB, PCA
PubMed
(). Noble metal targeting of thioredoxin reductase--covalent complexes with thioredoxin and thioredoxin-related protein of 14 kDa triggered by cisplatin. Free Radic Biol Med.
Applications
WB, IB, PCA
PubMed
(). Tautomycetin suppresses the TNFα/NF-κB pathway via inhibition of IKK activation. Int J Oncol
Applications
IP, Co-IP
PubMed
(). A feedback loop between the androgen receptor and a NEDD4-binding protein, PMEPA1, in prostate cancer cells. J Biol Chem.
Applications
WB, IB, PCA
PubMed

Frequently Asked Questions (13)

What is Rockland’s TrueBlot® product line?

Rockland’s TrueBlot® product line is designed to solve common experimental problems when performing immunoprecipitation/co-immunoprecipitation experiments and Western blots of immunoprecipitated samples (IP–Western blot). The product line consists of TrueBlot® monoclonal secondary antibodies and TrueBlot® IP beads.

What is the advantage of TrueBlot® secondaries over regular secondary antibodies?

TrueBlot® antibodies are specific to the whole IgG molecule and will not bind heavy or light chains. This is useful for binding target proteins with an expected MW near 25 kDa (light chains) and 50 kDa (heavy chains).

Why does it appear that the TrueBlot® antibody is binding heavy/light chains in my sample?

The sample must be fully reduced to eliminate cross-reactivity with heavy and light chains. Any reactivity to heavy and light chains could be due to incomplete reduction of your sample. Please be sure to optimize reducing conditions for your sample type.

How can I ensure my samples are fully reduced?

Samples can be fully reduced by heating at 90-100°C for 10 minutes in sample buffer containing reducing agent (for example, DTT to a final concentration of 50 mM or add β-Mercaptoethanol to a final concentration of 2% (v/v)).

With which species/isotypes are TrueBlot® secondary antibodies reactive?

TrueBlot® secondary antibodies are reactive with the IgG of their respective species. For example, if using a primary antibody raised in mouse (mouse host), use TrueBlot® anti-mouse Ig secondary antibodies for detection.

Do TrueBlot® secondary antibodies detect IgM?

TrueBlot® secondary antibodies have not been shown to detect IgM.

Do I need to preclear my lysate before the immunoprecipitation step?

Samples that have many other proteins present (such as lysates) may require preclearing to prevent interference in later IP and WB procedures. Recombinant protein samples require pre-clearing more often than serum samples.

How can I enrich/concentrate my lysate for a low expression of a protein of interest?

The immunoprecipitation procedure can be repeated several times to yield a more concentrated protein solution.

What is the recommended lysis buffer for TrueBlot® products?

The optimal lysis buffer will depend on your sample type. See protocol for buffer recommendations. Generally, 1X RIPA is used to lyse samples.

My target protein has the same MW as a heavy/light chain. How can I be sure that the band is the target protein and not a heavy/light chain?

Be sure to include a positive control for the primary antibody in your experiment and check that the sample is fully reduced.

What is the average size of TrueBlot® magnetic/agarose beads?

The beads are roughly 0.5 μm in diameter.

Why can’t I use Protein A or Protein G beads for the IP step in IP–Western blot when using rabbit species?

Using Protein A or Protein G beads with rabbit species results in contaminated bands. For best results when using rabbit species, use Rockland’s TrueBlot® Rabbit Ig IP beads, which are available in either a magnetic or agarose format. Please see individual TrueBlot® Rabbit product pages for additional details.

Can I use Protein A or Protein G beads for the IP step in IP–Western blot when using mouse, goat, or sheep species?

Yes. If you are using Protein A or Protein G beads with mouse, goat, or sheep species, we recommend the following: Determine the compatibility of Protein A or Protein G with your species and subtype, do not use excessive amounts of slurry, and choose the appropriate elution conditions. In some instances, harsher elution conditions can cause stripping of the subunits of Protein A or Protein G from the bead support and result in non-specific bands. For best results, we recommend using TrueBlot® Ig IP beads, which are available in either agarose or magnetic format.

Related Protocols

Adherent Cell Lysis Protocol
Bifunctional Cross-Linking Protocol
Biotin Binding Assay Protocol
Chromatin Immunoprecipitation (ChIP) Protocol
ELISA Protocol
Fluorescent Western Blotting Protocol
Histone Immunoblotting Protocol
Immunocytochemistry (ICC) Protocol
Immunofluorescence (IF) Protocol
Immunoprecipitation (IP) Protocol
In-Cell Western (ICW) Protocol
IP-WB with TrueBlot® Protocol
Multi-Lysate Western Blotting Protocol
Nuclear & Cytoplasmic Extract Protocol
Nucleosome ChIP Protocol
Peptide Competition Assay (PCA) Protocol
Re-ChIP Protocol
Sandwich ELISA Protocol for Collagen
Suspension Cultured Cell Lysis Protocol
Western Blotting (WB) Protocol

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Disclaimer

This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.

Related products to: Mouse TrueBlot® ULTRA: Anti-Mouse Ig HRP

Fluorescent TrueBlot®: Anti-Mouse Ig Fluorescein 18-0217-32
Sheep TrueBlot®: Anti-Sheep IgG HRP 18-8815-31
Goat TrueBlot®: Anti-Goat IgG HRP 18-8814-31
Rabbit TrueBlot®: Anti-Rabbit IgG HRP 18-8816-31
Fluorescent TrueBlot®: Anti-Rabbit IgG DyLight™ 800 18-4516-32

Related Resources

Selecting TrueBlot® Products

Literature
View Literature

IP-WB with TrueBlot® Protocol

Protocols
View Protocol

TrueBlot®: Publication Quality IP and Western Blot Images

Videos
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