ALP (Alkaline Phosphatase) BioAssay™ ELISA Kit (Rat)

Kits and Assays / Kits-ELISA, BioAssay™

4°C/-20°C

The Alkaline Phospatase (ALP) BioAssay™ ELISA Kit (Rat) is a quantitative sandwich assay for the detection of ALP in rat serum, plasma, tissue homogenates and other biological fluids.

Detectio Range:
0.625-40ng/ml

Sensitivity:
<0.375ng/ml

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

Assay Principle:
The microtiter plate provided in this kit has been pre-coated with an antibody specific to ALP. Standards and samples are added to the appropriate microtiter plate wells followed by a biotin-conjugated antibody specific to ALP. Streptavidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain ALP, Biotin-conjugated antibody and enzyme-conjugated Streptavidin complex 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 ALP in the sample is then determined by comparing the O.D. of the sample to the standard curve.

Kit Components:
*353845A: Microtiter Strips, 8x12 wells.
*353845B: Standard, 2 vials
353845C: Sample/Standard Dilution Buffer, 1x20ml
353845D: Antibody-Biotin (Concentrated), 1x120ul
353845E: Antibody Dilution Buffer, 1x10ml
353845F: Streptavidin-HRP Conjugate (SABC), 1x120ul
353845G: SABC Dilution Buffer, 1x10ml
353845H: TMB Substrate, 1x10ml
353845J: Stop Solution, 1x10ml
353845K: Wash Buffer (25X), 1x30ml

Storage and Stability:
Store unopened *353845A at 4ºC; store at -20ºC once opened. Store unopened *353845B 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, samples and control (zero) to wells!
2. Add 100ul standard or sample to each well and incubate for 90 minutes at 37°C
3. Add 100ul Antibody-Biotin working solution to each well and incubate for 60 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 plate 5 times.
7. Add 90ul TMB substrate. Incubate 15-30 minutes at 37°C
8. Add 50ul Stop Solution. Read at 450nm immediately.
9. Calculate results.