This kit is based on Double antibody-Sandwich ELISA detection method and takes 4h assay time. The microplate provided in this kit has been precoated with anti PINP antibody. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add biotinylated detection antibody. Then, it binds with PINP bound to precoated antibody. Wash unbound components and add HRP-Streptavidin Conjugate (SABC). Wash unbound components again and add TMB substrate solution. Then, TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. Calculate the concentration of PINP in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.
Manual
- Product Name
- Mouse PINP(N-terminal propeptide of Collagen alpha-1(I) chain) ELISA Kit
- Alias
- PINP ELISA Kit, P1NP ELISA Kit, N-Propeptide Of Type I Procollagen ELISA Kit, Procollagen I Amino Terminal Propeptide ELISA Kit
- Catalogue No.
- EM0481
- Size
- 48T/96T
- Species
- Mouse
- Sample Type
- Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples
- Detection Method
- Sandwich ELISA, Double Antibody
- Detection Wavelength
- OD450
- Reaction Duration
- 4 hours
- Range
- 78.125-5000pg/ml
- Sensitivity
- 46.875pg/ml
- Storage
- 2-8T)
Size(96T) Storage Condition for Opened Kit E001 ELISA Microplate(Dismountable) 8×6 8×12 Put the rest strips into a sealed foil bag with the desiccant. Stored for 1 month at 2-8 month at -20 Lyophilized Standard 1vial 2vial Put the rest standards into a desiccant bag. Stored for 1 month at 2-8 month at -20 Biotin-labeled Antibody(Concentrated, 100X) 60ul 120ul 2-8 HRP-Streptavidin Conjugate(SABC, 100X) 60ul 120ul E024 TMB Substrate 5ml 10ml E039 Sample Dilution Buffer 10ml 20ml 2-8 Antibody Dilution Buffer 5ml 10ml E049 SABC Dilution Buffer 5ml 10ml E026 Stop Solution 5ml 10ml E038 Wash Buffer(Concentrated, 25X) 15ml 30ml E006 Plate Sealer 3 pieces 5 pieces E007 Product Description 1 copy 1 copy - Required Instruments and Reagents
-
- Microplate reader (wavelength: 450nm)
- 37 incubator for cell culture is not recommenced.)
- Automated plate washer or multi-channel pipette/5ml pipettor (for manual washing purpose)
- Precision single (0.5-10μL, 5-50μL, 20-200μL, 200-1000μL) and multi-channel pipette with disposable tips(Calibration is required before use.)
- Sterile tubes and Eppendorf tubes with disposable tips
- Absorbent paper and loading slot
- Deionized or distilled water
- Assay Procedure Summary
-
- Step 1: Add 100ul standard or sample into each well, seal the plate and statically incubate for 90 minutes at 37: Add 100ul biotin-antibody working solution, seal the plate and statically incubate for 60 minutes at 37min each time.
- Step 3: Add 100ul HRP-Streptavidin Conjugate (SABC) working solution, seal the plate and statically incubate for 30 minutes at 37min each time.
- Step 4: Add 90ul TMB substrate solution, seal the plate and statically incubate for 10-20 minutes at 37: Add 50ul stop solution. Read at 450nm immediately and calculate.
- Standard Curve
-
This product is detected by QC department and meets performance required in the manual. (Laboratory Humidity: 20%-60%; Temperature: 18min at 37;margin-bottom: 0.5rem;">Due to different assay environments and operations, assay data below and standard curve are provided for reference. Experimenters should establish standard curve according to their own assay.
STD.(pg/ml) OD-1 OD-2 Average 0 0.086 0.088 0.087 78.125 0.195 0.201 0.198 156.25 0.298 0.306 0.302 312.5 0.483 0.497 0.49 625 0.907 0.933 0.92 1250 1.352 1.392 1.372 2500 1.91 1.966 1.938 5000 2.172 2.236 2.204 
- Recovery
-
Add a certain amount of PINP into the sample. Calculate the recovery by comparing the measured value with the expected amount of PINP in the sample.
Sample Type Recovery Range(%) Average(%) serum(n=10) 85-104 95 EDTA plasma(n=10) 94-103 99 Heparin plasma(n=10) 86-105 96 - Linearity
-
Dilute the sample with a certain amount of PINP at 1:2, 1:4 and 1:8 to get the recovery range.
Sample Type 1:2 1:4 1:8 serum(n=10) 85-103% 96-102% 86-104% EDTA plasma(n=10) 83-100% 86-99% 85-97% Heparin plasma(n=10) 82-90% 81-97% 81-95% - Precision(%)
-
Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on the same plate.
Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.
Item Intra-assay Precision Inter-assay Precision Sample 1 2 3 1 2 3 n 20 20 20 20 20 20 Mean (pg/ml) 162.1 616.8 2495 156.3 638.8 2518 Standard deviation 8.7 33.31 130.49 8.36 33.79 134.21 CV(%) 5.37 5.4 5.23 5.35 5.29 5.33 - Stability
-
Perform the stability test for the sealed kit at 37-8)
37 month 2-8 months Average(%) 80 95-100