Leptin (LEP) BioAssay ELISA Kit (Porcine)

£1,068.00 96 Tests

Leptin (LEP) BioAssay ELISA Kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of LEP in porcine serum, plasma, tissue homogenates and other biological fluids.

SKU: US026475 Categories: , ,

Additional information

Detection Range

31.2-2000pg/ml

Sensitivity

12.7pg/ml

Precision

Intra-Assay:
CV<10%

Inter-Assay:
CV<12%

Test Principle

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

Kit Components

*026475A: Microtiter Plate, 1×48 wells or 1×96 wells, Pre-coated; ready to use
*026475B: Standard, 2x1vial
026475C: Standard Diluent, 1x10ml or 1x20ml
*026475D: Detection Reagent A (green), 1x60ul or 1x120ul
*026475E: Detection Reagent B (red), 1x60ul or 1x120ul
026475F: Assay Diluent A, 1x6ml or 1x12ml
026475G: Assay Diluent B, 1x6ml or 1x12ml
026475H: TMB Substrate, 1×4.5ml or 1x9ml
*026475J: Positive Control, 246.2pg/ml, 1x1vial (Lyophilized)
026475K: Stop Solution, 1x3ml or 1x6ml
026475L: Wash Buffer, 30X, 1x10ml or 1x20ml

Precaution

The Stop Solution (026475K) suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.

Storage and Stability

Store *026475A, *026475B, *026475D, *026475E and *026475J at -20?C. After reconstitution of *026475J, store at 4?C and use within 5 days. Store all the other components at 4?C. Unused kit is stable for 6 months after receipt. Once kit components are opened, it is highly recommended to use remaining reagents within 1 month provided this is within the expiration date of the kit. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap.

Notes on 026475J

Positive Control

Intended Use

The control vials are from a porcine serum sample known to contain the target protein. It will provide a positive result in the ELISA assay, which indicates that the kit is optimized and working properly.

Characteristics

The control vial contains porcine LEP in porcine serum (lyophilized powder). Note: The porcine based serum used in this product is from a healthy specimen(s) of SPF standard (specific pathogen-free animals).

Expected Value

246.2pg/ml

Usage

Dissolve with 150ul standard diluent and let stand for 10min. Mix gently before use. In the experiment, add 100ul to an appropriate well of the plate and test its concentration along with standards and samples. Note: Upon receipt store Positive Control vial at -20?C. After reconstitution, store Positive Control vial at 4?C and use within 5 days.

Assay Procedure Summary

1. Prepare all reagents, samples and standards.
2. Add 100ul standard or sample to each well. Incubate 1 hour at 37?C.
3. Aspirate and add 100ul prepared Detection Reagent A. Incubate 1 hour at 37?C.
4. Aspirate and wash 3 times.
5. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37?C.
6. Aspirate and wash 5 times.
7. Add 90ul Substrate Solution. Incubate 10-20 minutes at 37?C.
8. Add 50ul Stop Solution. Read at 450nm immediately.

Shelf Life

6 Months