The Fibroblast Growth Factor 4 (FGF4) BioAssay™ ELISA Kit is a sandwich ELISA for in vitro quantitative measurement of FGF4 in mouse serum, plasma and other biological fluids.
Specificity:Recognizes mouse FGF4. No significant crossreactivity or interference between mouse FGF4 and analogues was observed.
Test Principle:This BioAssay™ ELISA kit uses Sandwich ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to mouse FGF4. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for mouse FGF4 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain mouse FGF4, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450nm ± 2nm. The OD value is proportional to the concentration of mouse FGF4. The concentration of mouse FGF4 in samples can be calculated by comparing the OD of the samples to the standard curve.
Precaution:The Stop Solution (382876K) included for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.
Storage and Stability:Store *382876A, *382876B, *382876D and *382876E at -20°C. Store all the other components at 4°C. Unused kit is stable for 6 months after receipt. 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 90 minutes at 37°C
3. Aspirate and add 100ul prepared Biotin Working Solution. Incubate 1 hour at 37°C
4. Aspirate and wash 3 times
5. Add 100ul prepared HRP Detection Working Solution. Incubate 30 minutes at 37°C
6. Aspirate and wash 5 times
7. Add 90ul Substrate. Incubate 15 minutes at 37°C
8. Add 50ul Stop Solution. Read at 450nm immediately.
