Complement Component 3a (C3a) BioAssay™ ELISA Kit (Human)

Blue Ice

38220000

The Complement Component 3a (C3a) BioAssay™ ELISA Kit (Human) is a sandwich enzyme immunoassay for the in vitro quantitative measurement of C3a in human serum, plasma, tissue homogenates, cell lysates, cell culture supernatants and other biological fluids.


Detection Range:
78-5, 000pg/ml

Sensitivity:
<30pg/ml

Precision:
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Test Principle:
The microtiter plate provided in this kit has been pre-coated with an antibody specific to C3a. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to C3a. 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 C3a, 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 C3a in the sample is then determined by comparing the O.D. of the sample to the standard curve.

Kit Components:
*024345A: Microtiter Plate, 1x96 wells. Pre-coated; ready to use.
*024345B: Standard, 2x1vial
024345C: Standard Diluent, 1x20ml
*024345D: Detection Reagent A, 1x120ul
*024345E: Detection Reagent B, 1x120ul
024345F: Assay Diluent A, 1x12ml
024345G: Assay Diluent B, 1x12ml
024345H: TMB Substrate, 1x9ml
024345K: Stop Solution, 1x6ml
024345L: Wash Buffer, 30X, 1x20ml

Storage and Stability:
Store *024345A, *024345B, *024345D and *024345E 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 for 1 hour at 37°C.
3. Aspirate and 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 TMB Substrate. Incubate 10-20 minutes at 37°C.
8. Add 50ul Stop Solution. Read at 450nm immediately.