Lipoxin A4 (LXA4) BioAssay™ ELISA Kit (Universal)

Kits and Assays / Kits-ELISA (Eicosanoids and Eicosanoid Metabolism), BioAssay™

4°C/-20°C

The Lipoxin A4 (LXA4) BioAssay™ ELISA Kit is a quantitative competitive assay for the detection of LXA4 in serum, plasma, tissue homogenates and other biological fluids.

Detection Range:
0.781-50ng/ml

Sensitivity:
<0.469ng/ml

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

Assay Principle:
The microtiter plate provided in this kit has been pre-coated with LXA4. LXA4 in the standard or sample competes with a fixed amount of plate-coated LXA4 for antibody binding sites on the Antibody-Biotin reagent. Excess conjugate and unbound sample or standard are washed from the plate, after which Streptavidin-HRP (SABC) is added to each 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 LXA4 in the samples is determined by an inverse correlation between LXA4 in the sample and the assay signal intensity.

Kit Components:
*357160A: Microtiter Strips, 1x96 wells (8x12 wells)
*357160B: Standard, 2x1 vials
357160C: Sample/Standard Dilution Buffer, 1x20ml
357160D: Antibody-Biotin Concentrate, 1x60ul
357160E: Antibody Dilution Buffer, 1x10ml
357160F: Streptavidin-HRP Conjugate (SABC), 1x120ul
357160G: SABC Dilution Buffer, 1x10ml
357160H: TMB Substrate, 1x10ml
357160J: Stop Solution, 1x10ml
357160K: Wash Buffer, 25X, 1x30ml

Storage and Stability:
Store unopened *357160A at 4ºC; store at -20ºC once opened. Store unopened *357160B 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. Immediately 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.