The Human Granulocyte Colony Stimulating Factor (G-CSF) ELISA Kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of G-CSF in human serum, plasma other biological fluids.
Detection Range
15.6-1,000pg/ml
Test Principle
The microtiter plate provided in this kit has been pre-coated with an antibody specific to human G-CSF. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for human G-CSF. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain G-CSF, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulfuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of G-CSF in the sample is then determined by comparing the O.D. of the sample to the standard curve.
Kit Components
*024323A: Microtiter Plate, 1x96 wells, Pre-coated; ready to use *024323B: Standard, 2x1vial 024323C: Standard Diluent, 1x20ml *024323D: Detection Reagent A, 1x120ul *024323E: Detection Reagent B, 1x120ul 024323F: Assay Diluent A (2x),1x6ml 024323G: Assay Diluent B (2x), 1x6ml 024323H: TMB Substrate, 1x9ml 024323K: Stop Solution, 1x6ml 024323L: Wash Buffer, 30X, 1x20ml
Storage and Stability
Store *024323A, *024323B, *024323D and *024323E at -20°C. Store all the other components at 4°C. Unused kit is stable for 6 months. Once kit components are opened, it is highly recommended to use remaining reagents within 1 month provided this is within the expiration date of the kit. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap.
Assay Procedure Summary
1. Prepare all reagents, samples and standards. 2. Add 100ul standard or sample to each well. Incubate 2 hours at 37°C. 3. Add 100ul prepared Detection Reagent A. Incubate 1 hour at 37°C. 4. Aspirate and wash 3 times. 5. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37°C. 6. Aspirate and wash 5 times. 7. Add 90ul 024323H: TMB Substrate. Incubate 15-25 minutes at 37°C. 8. Add 50ul 024323K: Stop Solution. Read at 450nm immediately.