Platelet Activating Factor (PAF) BioAssay™ ELISA Kit (General)

The Platelet Activating Factor (PAF) BioAssay™ ELISA Kit is a quantitative competitive assay for the detection of PAF in serum, plasma, tissue homogenates and other biological fluids.

1.56-100ng/ml

<0.938ng/ml

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

The microtiter plate provided in this kit has been pre-coated with PAF. PAF in the standard or sample competes with a fixed amount of plate-coated PAF 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 PAF in the samples is determined by comparing the absorbance of the samples to the standard curve.

*396541A: Microtiter Strips, 8x12 wells.
*396541B: Standard, 2 x1vial
396541C: Sample/Standard Dilution Buffer, 1x20ml
396541D: Antibody-Biotin Concentrate, 1x60ul
396541E: Antibody Dilution Buffer, 1x10ml
396541F: Streptavidin-HRP Conjugate (SABC), 1x120ul
396541G: SABC Dilution Buffer, 1x10ml
396541H: TMB Substrate, 1x10ml
396541J: Stop Solution, 1x10ml
396541K: Wash Buffer, 25X, 1x30ml

Store unopened *396541A at 4ºC; store at -20ºC once opened. Store unopened *396541B 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.

1. Wash plate 2 times before adding standards, samples and control (zero) 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.

This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.