Homocysteine (HCY) BioAssay™ ELISA Kit (Mouse)

Kits and Assays / Kits-ELISA, BioAssay™

4°C/-20°C

Homocysteine (HCY) BioAssay™ ELISA Kit is a quantitative competitive assay for the detection of HCY in mouse serum, plasma, tissue homogenates and other biological fluids.

Detection Range:
7.81-500pmol/ml

Sensitivity:
<4.69pmol/ml

Precision:
Intra-Assay CV: <8%
Inter-Assay CV: <10%

Assay Principle:
The microtiter plate provided in this kit has been pre-coated with HCY. HCY in the standard or sample competes with a fixed amount of plate-coated HCY for antibody binding sites on the Antibody-Biotin reagent. Excess conjugate and unbound sample or standard are washed from the plate, after which Streptavidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. TMB substrate is then added to each well. The enzyme-substrate reaction is terminated by the addition of a sulfuric acid solution and the resulting color is measured spectrophotometrically at 450nm. The concentration of HCY in the samples is determined by an inverse correlation between HCY in the sample and the assay signal intensity.

Kit Components:
*356228A: Microtiter Strips, 1x48 wells or 1x96 wells.
*356228B: Standard, 1x1vial or 2x1 vial
356228C: Sample/Standard Dilution Buffer, 1x10ml or 1x20ml
356228D: Antibody-Biotin (Concentrated), 1x30ul or 1x60ul
356228E: Antibody Dilution Buffer, 1x5ml or 1x10ml
356228F: Streptavidin (HRP) (SABC, 1x60ul or 1x120ul
356228G: SABC Dilution Buffer, 1x5ml or 1x10ml
356228H: TMB Substrate, 1x5ml or 1x10ml
356228J: Stop Solution, 1x5ml or 1x10ml
356228K: Wash Buffer, 25X, 1x15ml or 1x30ml

Storage and Stability:
Store unopened *356228A at 4ºC; store at -20ºC once opened. Store unopened *356228B at 4°C; once reconstituted, store at 4°C for up to 12 hours or at -20°C for up to 48 hours. Store other components at 4°C. Kit is stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap.

Assay Summary:
1. Wash plate 2 times before adding standards and samples to wells!
2. Add 50ul standard or sample to each well.
3. Add 50ul Antibody (Biotin) working solution to each well and incubate for 45 minutes at 37°C
4. Aspirate and wash 3 times.
5. Add 100ul SABC working solution to each well. Incubate for 30 minutes at 37°C
6. Aspirate and wash 5 times.
7. Add 90ul TMB substrate. Incubate 15-20 minutes at 37°C
8. Add 50ul Stop Solution. Read at 450nm immediately.
9. Calculate results