The COL18a1 (Collagen Type XVIII Alpha 1) BioAssay™ ELISA Kit (Human) is a quantitative sandwich ELISA for the detection of COL18alpha1 in human serum, plasma and other biological fluids
Specificity:Recognizes natural and some recombinant Human COL18a1. No significant cross-reactivity or interference between Human COL18a1 and analogues was observed.
Test Principle:The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to human COL18alpha1 (Collagen Type XVIII Alpha 1). 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 human COL18alpha1 (Collagen Type XVIII Alpha 1) 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 human COL18alpha1 (Collagen Type XVIII Alpha 1), 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 450 nm ± 2 nm. The OD value is proportional to the concentration of human COL18alpha1 (Collagen Type XVIII Alpha 1). The concentration of human COL18alpha1 (Collagen Type XVIII Alpha 1) in samples can be calculated by comparing the OD of the samples to the standard curve.
Storage and Stability:Unused kit is stable for 6 months after receipt. Store Microtiter Strips, Standard, and Antibody-Biotin at 4°C if kit will be used within 30 days; otherwise store at -20°C until expiration date. Store the rest of the components at 4°C. Protect HRP Conjugate and Substrate Reagent from light. For maximum recovery, 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.
