Human Leptin ELISA Kit

Catalog No.: BEK1136

Size: 96T

Range: 62.5 pg/ml-4000 pg/ml

Sensitivity < 5 pg/ml

Storage and Expiration: Store at 2-8℃ for 6 months, or at -20℃ for 12 months. 

Application: For quantitative detection of Leptin in human serum, plasma, body fluids, tissue lysates or cell culture supernates. 

Introduction

Leptin is a 16 kDa protein hormone that in humans encoded by the Ob gene, the gene is located on chromosome 7 in humans. The Leptin is one of the most important adipose derived hormones. It is manufactured primarily in the adipocytes of white adipose tissue, and the level of circulating leptin is proportional to the total amount of fat in the body. It plays a critical role in the regulation of body weight by inhibiting food intake and stimulating energy expenditure. In addition to its effects on body weight, leptin has a variety of other functions, including the regulation of hematopoiesis, angiogenesis, wound healing, and the immune and inflammatory response. In the clinical, Leptin has traditionally been regarded as a link between fat mass, food intake, and energy expenditure.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-Leptin polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-Leptin polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the Leptin amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of Leptin can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-Human Leptin antibody
2. Lyophilized Leptin standards: 2 tubes (10ng / tube)
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Human Leptin antibody (Concentrated): 130μl. Dilution: 1:100
5. Antibody diluent buffer: 12ml
6. Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130μl. Dilution: 1:100
7. ABC diluent buffer: 12ml
8. TMB substrate: 10ml
9. Stop solution: 10ml
10. Wash buffer (25X): 30ml

Note: Reconstitute standards and test samples with Kit Component 3.

Material Required But Not Provided

1. 37℃ incubator
2. Microplate reader (wavelength: 450nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5ml of Eppendorf tubes
7. Plate cover
8. Absorbent filter papers
9. Plastic or glass container with volume of above 1L