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> Blocking Buffer (2X) for Fluorescent Western Blotting

Blocking Buffer (2X) for Fluorescent Western Blotting

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Cat# MB-070-0050

Size : 50mL

Brand : Rockland Immunochemicals

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Specifications for Blocking Buffer (2X) for Fluorescent Western Blotting

Product Details

Blocking Buffer (2X) for Fluorescent Western Blotting - MB-070-0050
Multiplex Blocking Buffer, Fluorescent Blocking Buffer, Blocking Solution, Blocking Buffer Western Blot, IRDye Western Blot Blocking Buffer, Alexa Dye Blocking Buffer, DyLight Blocking Buffer

Target Details

Blocking buffer is specifically formulated to achieve superior reproducible Western blotting images using this system.

Application Details

WB
Cellular Assay, ELISA, IF, IHC, Microarray, Other  - View References
This product is a 2X concentrated stock solution. Prepare a 1X working solution by diluting 1 part 2X concentrate with 1 parts TBS or equivalent. This buffer was meticulously prepared using ultra pure reagents dissolved in pharmaceutical grade water (WFI) and consists of a proprietary protein formulation in TRIS buffered saline at pH 7.6 with thimerosal added as an antimicrobial agent.

Formulation

2X
See application note.
Thimerosal is added as an antimicrobial agent.

Shipping & Handling

Wet Ice
Store blocking buffer at 4° C prior to opening. DO NOT FREEZE.
Expiration date is six (6) months from date of receipt.

Background

This blocking buffer is ideal for infrared Western blotting. Fluorescence technology is widely used to detect proteins. However, many common visible fluorophores often result in considerable background fluorescence in the visible range. Visible fluorophores are rarely used for membrane-based protein detection because of this high background. IRDye™800, IRDye™700DX, Alexa Fluor® 680 and Cy5.5™ antibody and reagent conjugates are specifically designed for protein detection methods that use longer-wavelength, near-infrared (IR) fluorophores to visualize proteins in western blotting and other applications. IRDye™800 and IRDye™700DX fluoresce outside the range of most autofluorescence and therefore specific signal is sharply evident from any background giving the best possible signal-to-noise ratio. Detection levels in the picogram range on Western blots rival the sensitivity of chemiluminescence on film. IRDye™800 conjugates are also suitable for immunofluorescence microscopy using commercially available excitation/emission filters in the 780nm/820nm range.  Dual simultaneous labeling in Western blots or microscopy is achieved when IRDye™800 conjugates are used in conjunction with IRDye™700 or Cy5.5™ conjugates. IRDye™800 and IRDye™700DX conjugates provide an ultra-sensitive and convenient alternative to standard chemiluminescent protein detection methods, as well as a valuable tool for multicolor imaging. Once reacted with the membrane and dried, IRDye™800 and IRDye™700DX conjugated antibody-protein complexes are very stable, and membranes can be stored protected from light, re-washed and/or rescanned.

References (102)

(). Epitopes of Naturally Acquired and Vaccine-Induced Anti-Ebola Virus Glycoprotein Antibodies in Single Amino Acid Resolution. Biotechnol J.
Applications
Microarray
PubMed
(). Versicolorin A, a precursor in aflatoxins biosynthesis, is a food contaminant toxic for human intestinal cells. Environ Int.
Applications
WB, IB, PCA
PubMed
(). Expression and Function of Mas-Related G Protein-Coupled Receptor D and Its Ligand Alamandine in Retina. Mol Neurobiol.
Applications
WB, IB, PCA
PubMed
(). A proteolytic C-terminal fragment of Nogo-A (reticulon-4A) is released in exosomes and potently inhibits axon regeneration. J Biol Chem.
Applications
WB, IB, PCA
PubMed
(). The stress-responsive gene GDPGP1/mcp-1 regulates neuronal glycogen metabolism and survival. J Cell Biol.
Applications
WB, IB, PCA
PubMed
(). Pre-clinical characterisation of E2814, a high-affinity antibody targeting the microtubule-binding repeat domain of tau for passive immunotherapy in Alzheimer's disease. Acta Neuropathol Commun.
Applications
Microarray
PubMed
(). Immunization with full-length Plasmodium falciparum merozoite surface protein 1 is safe and elicits functional cytophilic antibodies in a randomized first-in-human trial. NPJ Vaccines.
Applications
Microarray
PubMed
(). Human HMGN1 and HMGN2 are not required for transcription-coupled DNA repair. Sci Rep.
Applications
WB, IB, PCA
PubMed
(). On-Chip Neo-Glycopeptide Synthesis for Multivalent Glycan Presentation. Chemistry.
Applications
In Vitro Binding Assay
PubMed
(). The cooperative action of CSB, CSA, and UVSSA target TFIIH to DNA damage-stalled RNA polymerase II. Nat Commun.
Applications
WB, IB, PCA
PubMed
(). TrkB agonists prevent postischemic emergence of refractory neonatal seizures in mice. JCI Insight.
Applications
WB, IB, PCA
PubMed
(). A cross-reactive mouse monoclonal antibody against rhinovirus mediates phagocytosis in vitro. Sci Rep.
Applications
Microarray
PubMed
(). Hypoxia regulates the degradation of non-nuclear organelles during lens differentiation through activation of HIF1a Exp Eye Res.
Applications
WB, IB, PCA
PubMed
(). Expression of different L1 isoforms of Mastomys natalensis papillomavirus as mechanism to circumvent adaptive immunity. Elife.
Applications
Microarray
PubMed
(). Identification of truncated C-terminal fragments of the Alzheimer's disease amyloid protein precursor derived from sequential proteolytic pathways. J Neurochem.
Applications
WB, IB, PCA
PubMed
(). Small-molecule inhibition of Lats kinases promotes Yap-dependent proliferation in postmitotic mammalian tissues. bioRxiv
Applications
WB, IB, PCA
Source
(). Cardiac Cell-Derived Matrices Impart Age-Specific Functional Properties to Human Cardiomyocytes. bioRxiv
Applications
WB, IB, PCA
Source
(). Trem2 Deletion Reduces Late-Stage Amyloid Plaque Accumulation, Elevates the Aβ42:Aβ40 Ratio, and Exacerbates Axonal Dystrophy and Dendritic Spine Loss in the PS2APP Alzheimer's Mouse Model. J Neurosci.
Applications
Other
PubMed
(). SNX27 regulates DRA activity and mediates its direct recycling by PDZ-interaction in early endosomes at the apical pole of Caco2 cells. Am J Physiol Gastrointest Liver Physiol.
Applications
WB, IB, PCA
PubMed
(). Organic dust induced mitochondrial dysfunction could be targeted via cGAS-STING or mitochondrial NOX-2 inhibition. bioRxiv
Applications
WB, IB, PCA
Source
(). In vivo proximity proteomics of nascent synapses reveals a novel regulator of cytoskeleton-mediated synaptic maturation. Nature Communications
Applications
IF, Confocal Microscopy; IHC, ICC, Histology
PubMed
(). Amplified Host Defense by Toll-Like Receptor-Mediated Downregulation of the Glucocorticoid-Induced Leucine Zipper (GILZ) in Macrophages. Frontiers in Immunology
Applications
WB, IB, PCA
PubMed
(). Alzheimer's Disease Risk Factor Pyk2 Mediates Amyloid-β-Induced Synaptic Dysfunction and Loss. J Neurosci.
Applications
WB, IB, PCA
PubMed
(). Lipidomic Analysis of α-Synuclein Neurotoxicity Identifies Stearoyl CoA Desaturase as a Target for Parkinson Treatment. Mol Cell.
Applications
WB, IB, PCA
PubMed
(). Maelstrom Represses Canonical Polymerase II Transcription within Bi-directional piRNA Clusters in Drosophila melanogaster. Mol Cell.
Applications
WB, IB, PCA
PubMed
(). Pyk2 Signaling through Graf1 and RhoA GTPase Is Required for Amyloid-β Oligomer-Triggered Synapse Loss. J Neurosci.
Applications
WB, IB, PCA
PubMed
(). Rescue of Transgenic Alzheimer's Pathophysiology by Polymeric Cellular Prion Protein Antagonists. Cell Rep.
Applications
WB, IB, PCA
PubMed
(). Plexina2 and CRMP2 Signaling Complex Is Activated by Nogo-A-Liganded Ngr1 to Restrict Corticospinal Axon Sprouting after Trauma. J Neurosci.
Applications
WB, IB, PCA
PubMed
(). High-throughput epitope profiling of antibodies in the plasma of Alzheimer's disease patients using random peptide microarrays. Sci Rep.
Applications
Microarray
PubMed
(). Anti-PrPC antibody rescues cognition and synapses in transgenic alzheimer mice. Ann Clin Transl Neurol.
Applications
WB, IB, PCA
PubMed
(). Immunization of cats to induce neutralizing antibodies against Fel d 1, the major feline allergen in human subjects. J Allergy Clin Immunol.
Applications
Microarray
PubMed
(). The RNA-Binding ATPase, Armitage, Couples piRNA Amplification in Nuage to Phased piRNA Production on Mitochondria. Mol Cell.
Applications
WB, IB, PCA
PubMed
(). A Monoclonal Antibody to M-Phase Phosphoprotein 1/Kinesin-Like Protein KIF20B. Monoclon Antib Immunodiagn Immunother.
Applications
Microarray
PubMed
(). Toll-Like Receptor 2 Release by Macrophages: An Anti-inflammatory Program Induced by Glucocorticoids and Lipopolysaccharide. Front Immunol.
Applications
WB, IB, PCA
PubMed
(). Mitochondrial calcium exchange links metabolism with the epigenome to control cellular differentiation. Nat Commun.
Applications
WB, IB, PCA
PubMed
(). Biphasic Response of Protein Kinase A to Cyclic Adenosine Monophosphate Triggers Distinct Epithelial Phenotypes. bioRxiv
Applications
WB, IB, PCA
Source
(). MICU1 regulates mitochondrial cristae structure and function independent of the mitochondrial calcium uniporter channel. bioRxiv
Applications
WB, IB, PCA
Source
(). mitoTev-TALE: a monomeric DNA editing enzyme to reduce mutant mitochondrial DNA levels. EMBO Molecular Medicine
Applications
WB, IB, PCA
PubMed
(). Identification of the antigenic epitopes of maternal autoantibodies in autism spectrum disorders. Brain Behav Immun.
Applications
Microarray
PubMed
(). The nociceptin receptor inhibits axonal regeneration and recovery from spinal cord injury. Sci Signal.
Applications
WB, IB, PCA
PubMed
(). Protein Corona in Response to Flow: Effect on Protein Concentration and Structure. Biophys J.
Applications
WB, IB, PCA
PubMed
(). Mycobacterium tuberculosis-specific CD4+ and CD8+ T cells differ in their capacity to recognize infected macrophages. PLoS Pathog.
Applications
WB, IB, PCA
PubMed
(). BNIP3L/NIX is required for elimination of mitochondria, endoplasmic reticulum and Golgi apparatus during eye lens organelle-free zone formation. Exp Eye Res.
Applications
WB, IB, PCA
PubMed
(). Combinatorial Synthesis of Macromolecular Arrays by Microchannel Cantilever Spotting (µCS). Adv Mater.
Applications
Microarray
PubMed
(). Thrombospondin receptor α2δ-1 promotes synaptogenesis and spinogenesis via postsynaptic Rac1. J Cell Biol.
Applications
WB, IB, PCA
PubMed
(). A Single Mechanism of Biogenesis, Initiated and Directed by PIWI Proteins, Explains piRNA Production in Most Animals. Mol Cell.
Applications
WB, IB, PCA
PubMed
(). Generation and characterization of monoclonal antibodies that recognize human and murine supervillin protein isoforms. PLoS One.
Applications
Microarray
PubMed
(). Liquid and Hydrogel Phases of PrPC Linked to Conformation Shifts and Triggered by Alzheimer's Amyloid-β Oligomers. Mol Cell.
Applications
WB, IB, PCA
PubMed
(). KDEL Receptors Are Differentially Regulated to Maintain the ER Proteome under Calcium Deficiency. Cell Rep.
Applications
WB, IB, PCA
PubMed
(). Development of Neutralizing and Non-neutralizing Antibodies Targeting Known and Novel Epitopes of TcdB of Clostridioides difficile. Front Microbiol.
Applications
Microarray
PubMed
(). Interaction of the Human Respiratory Syncytial Virus matrix protein with cellular adaptor protein complex 3 plays a critical role in trafficking. PLOS One
Applications
WB, IB, PCA
PubMed
(). Early Activation of Experience-Independent Dendritic Spine Turnover in a Mouse Model of Alzheimer's Disease. Cereb Cortex.
Applications
WB, IB, PCA
PubMed
(). Analysis of Cohesin Function in Gene Regulation and Chromatin Organization in Interphase. Methods Mol Biol.
Applications
WB, IB, PCA
PubMed
(). Opposing effects of progranulin deficiency on amyloid and tau pathologies via microglial TYROBP network. Acta Neuropathol.
Applications
WB, IB, PCA
PubMed
(). Induction of Immune Tolerance to Foreign Protein via Adeno-Associated Viral Vector Gene Transfer in Mid-Gestation Fetal Sheep. PLoS One.
Applications
E, EIA
PubMed
(). A biomimetic gelatin-based platform elicits a pro-differentiation effect on podocytes through mechanotransduction. Sci Rep.
Applications
WB, IB, PCA
PubMed
(). Lifespan differences between queens and workers are not explained by rates of molecular damage. Exp Gerontol.
Applications
WB, IB, PCA
PubMed
(). The C Terminus of the Herpes Simplex Virus UL25 Protein Is Required for Release of Viral Genomes from Capsids Bound to Nuclear Pores. J Virol.
Applications
WB, IB, PCA
PubMed
(). Nanoparticle-induced oxidation of corona proteins initiates an oxidative stress response in cells. Nanoscale.
Applications
WB, IB, PCA
PubMed
(). Loss of TMEM106B Ameliorates Lysosomal and Frontotemporal Dementia-Related Phenotypes in Progranulin-Deficient Mice. Neuron.
Applications
WB, IB, PCA
PubMed
(). Systematic and Quantitative Assessment of Hydrogen Peroxide Reactivity With Cysteines Across Human Proteomes. Mol Cell Proteomics.
Applications
WB, IB, PCA
PubMed
(). Conditional Deletion of Prnp Rescues Behavioral and Synaptic Deficits after Disease Onset in Transgenic Alzheimer's Disease. J Neurosci.
Applications
WB, IB, PCA
PubMed
(). Identification of two conserved B-cell epitopes in the gp90 of reticuloendothelial virus using peptide microarray. Vet Microbiol.
Applications
Microarray
PubMed
(). Mapping Putative B-Cell Zika Virus NS1 Epitopes Provides Molecular Basis for Anti-NS1 Antibody Discrimination between Zika and Dengue Viruses. ACS Omega.
Applications
Microarray
PubMed
(). Metabotropic glutamate receptor 5 couples cellular prion protein to intracellular signalling in Alzheimer's disease. Brain.
Applications
WB, IB, PCA
PubMed
(). Serum peptide reactivities may distinguish neuromyelitis optica subgroups and multiple sclerosis. Neurol Neuroimmunol Neuroinflamm.
Applications
Microarray
PubMed
(). Identification of a Highly Conserved Epitope on Avian Influenza Virus Non-Structural Protein 1 Using a Peptide Microarray. PLoS One.
Applications
Microarray
PubMed
(). Assembly Dynamics and Stoichiometry of the Apoptosis Signal-regulating Kinase (ASK) Signalosome in Response to Electrophile Stress. Mol Cell Proteomics.
Applications
WB, IB, PCA
PubMed
(). High-Density Peptide Arrays for Malaria Vaccine Development. Methods Mol Biol.
Applications
WB, IB, PCA
PubMed
(). Properties of Two Enterovirus Antibodies that are Utilized in Diabetes Research. Sci Rep.
Applications
Microarray
PubMed
(). High-flexibility combinatorial peptide synthesis with laser-based transfer of monomers in solid matrix material. Nat Commun.
Applications
Other
PubMed
(). Oligomers of Amyloid β Prevent Physiological Activation of the Cellular Prion Protein-Metabotropic Glutamate Receptor 5 Complex by Glutamate in Alzheimer Disease. J Biol Chem.
Applications
WB, IB, PCA
PubMed
(). A single amino acid substitution alter antigenicity of Glycosylated protein 4 of HP-PRRSV. Virol J.
Applications
Microarray
PubMed
(). Transient Hepatic Overexpression of Insulin-Like Growth Factor 2 Induces Free Cholesterol and Lipid Droplet Formation. Front Physiol.
Applications
WB, IB, PCA
PubMed
(). Increased Intimal Hyperplasia After Vascular Injury in Male Androgen Receptor-Deficient Mice. Endocrinology.
Applications
WB, IB, PCA
PubMed
(). Identification of novel antigens contributing to autoimmunity in cardiovascular diseases. Clin Immunol.
Applications
Microarray
PubMed
(). Validation of Cross-Species Reactivity of the VEGF-A/PDGFRβ Bifunctional Antibody PF-06653157. J Ocul Pharmacol Ther.
Applications
WB, IB, PCA
PubMed
(). Dynamic Phosphorylation of Apoptosis Signal Regulating Kinase 1 (ASK1) in Response to Oxidative and Electrophilic Stress. Chem Res Toxicol.
Applications
WB, IB, PCA
PubMed
(). TiO2 Nanoparticles Alter the Expression of Peroxiredoxin Antioxidant Genes. J. Phys. Chem. C
Applications
WB, IB, PCA
Source
(). Printed peptide arrays identify prognostic TNC serumantibodies in glioblastoma patients. Oncotarget
Applications
Microarray
PubMed
(). Proteomic analysis of physiological versus pathological cardiac remodeling in animal models expressing mutations in myosin essential light chains. Journal of Muscle Research and Cell Motility
Applications
WB, IB, PCA
PubMed
(). Pattern recognition receptors in the gut: analysis of their expression along the intestinal tract and the crypt/villus axis. Physiol Rep.
Applications
WB, IB, PCA
PubMed
(). Spot peptide arrays and SPR measurements: throughput and quantification in antibody selectivity studies. J Mol Recognit.
Applications
Microarray
PubMed
(). Cross-Talk between Carbon Metabolism and the DNA Damage Response in S. cerevisiae. Cell Rep.
Applications
WB, IB, PCA
PubMed
(). Huntingtin is required for normal excitatory synapse development in cortical and striatal circuits. J Neurosci.
Applications
WB, IB, PCA
PubMed
(). Lysosome size, motility and stress response regulated by fronto-temporal dementia modifier TMEM106B. Mol Cell Neurosci.
Applications
WB, IB, PCA
PubMed
(). Isolation and characterization of the herpes simplex virus 1 terminase complex. J Virol.
Applications
WB, IB, PCA
PubMed
(). Analysis of autoantibodies to 3-hydroxy-3-methylglutaryl-coenzyme A reductase using different technologies. J Immunol Res.
Applications
Microarray
PubMed
(). Antigenic characteristics of glycosylated protein 3 of highly pathogenic porcine reproductive and respiratory syndrome virus. Virus Res.
Applications
Microarray
PubMed
(). Redox processes inform multivariate transdifferentiation trajectories associated with TGFβ-induced epithelial-mesenchymal transition. Free Radic Biol Med.
Applications
Other
PubMed
(). Therapeutic molecules and endogenous ligands regulate the interaction between brain cellular prion protein (PrPC) and metabotropic glutamate receptor 5 (mGluR5). J Biol Chem.
Applications
WB, IB, PCA
PubMed
(). Linear epitope recognition antibodies strongly respond to the C-terminal domain of HP-PRRSV GP5. Vet Microbiol.
Applications
Microarray
PubMed
(). Autophagy-dependent PELI3 degradation inhibits proinflammatory IL1B expression. Autophagy.
Applications
WB, IB, PCA
PubMed
(). Astrocytes refine cortical connectivity at dendritic spines. Elife.
Applications
WB, IB, PCA
PubMed
(). SUMOylation regulates the SNF1 protein kinase. Proc Natl Acad Sci U S A.
Applications
WB, IB, PCA
PubMed
(). WhiB7, an Fe-S-dependent transcription factor that activates species-specific repertoires of drug resistance determinants in actinobacteria. J Biol Chem.
Applications
WB, IB, PCA
PubMed
(). Mutational analysis of sclerostin shows importance of the flexible loop and the cystine-knot for Wnt-signaling inhibition. PLoS One.
Applications
Cellular assay
PubMed
(). Alzheimer amyloid-β oligomer bound to postsynaptic prion protein activates Fyn to impair neurons. Nat Neurosci.
Applications
WB, IB, PCA
PubMed
(). Sensing immune responses with customized peptide microarrays. Biointerphases.
Applications
Microarray
PubMed
(). Residues of the UL25 protein of herpes simplex virus that are required for its stable interaction with capsids. J Virol.
Applications
WB, IB, PCA
PubMed
(). Endo-lysosomal vesicles positive for Rab7 and LAMP1 are terminal vesicles for the transport of dextran. PLoS One.
Applications
WB, IB, PCA
PubMed
(). Coordinated regulation of extracellular matrix synthesis by the microRNA-29 family in the trabecular meshwork. Invest Ophthalmol Vis Sci.
Applications
WB, IB, PCA
PubMed

Related Protocols

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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.

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