Human Adiponectin/Acrp 30 ELISA Kit

Catalog No.: BEK1005

Size: 96T

Range: 780 pg/ml - 50,000 pg/ml

Sensitivity < 65 pg/ml

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

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

Introduction

Adiponectin (also referred to as GBP-28, apM1, AdipoQ and Acrp30) is a protein which in humans is encoded by the ADIPOQ gene. This gene spans 16 kb, contains 3 exons, and maps to to chromosome 3q27. Adiponectin is a 244-amino-acid-long polypeptide, and it has four distinct regions. It is a hormone secreted by adipocytes (and also from the placenta in pregnancy) that regulates energy homeostasis and glucose and lipid metabolism. Adipocytes also produce and secrete proteins such as leptin, adipsin, various other complement components and tumor necrosis factor, suggesting a possible link to the immune system.

Principle of the Assay 

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-Adiponectin polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-Adiponectin 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 Adiponectin amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of Adiponectin can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-human Adiponectin antibody
2. Lyophilized human Adiponectin standards: 2 tubes (100ng / tube)
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-human Adiponectin 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