Leptin (LEP) BioAssay™ ELISA Kit (Rat)

Kits and Assays / Kits-ELISA (Obesity), BioAssay™

4°C/-20°C

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

Detection Range:
0.156-10ng/ml

Sensitivity:
<0.094ng/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 Leptin. Standards and samples are added to the appropriate microtiter plate wells followed by a biotin-conjugated antibody specific to Leptin. 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 Leptin, 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 Leptin in the sample is then determined by comparing the O.D. of the sample to the standard curve.

Kit Components:
357025A: Microtiter Strips, 8x12 wells.
357025B: Standard, 2 vials
357025C: Sample/Standard Dilution Buffer, 1x20ml
357025D: Antibody-Biotin (Concentrated), 1x120ul
357025E: Antibody Dilution Buffer, 1x10ml
357025F: Streptavidin-HRP Conjugate (SABC), 1x120ul
357025G: SABC Dilution Buffer, 1x10ml
357025H: TMB Substrate, 1x10ml
357025J: Stop Solution, 1x10ml
357025K: Wash Buffer (25X), 1x30ml

Storage and Stability:
Store unopened 357025A at 4ºC; store at -20ºC once opened. Store unopened357025B at 4°C; once reconstituted store at 4°C for up to 12 hours or at -20°C for up to 48 hours. Store 357025F and 357025H protected from light at 4°C. Store all 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. Aspirate the liquid and wash plate 2 times.
3. Add 100ul Antibody-Biotin working solution to each well and incubate for 60 minutes at 37°C
4. Aspirate the liquid and wash plate 3 times.
5. Add 100ul SABC working solution to each well. Incubate for 30 minutes at 37°C
6. Aspirate the liquid 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.