Monkey Adiponectin, ADP ELISA Kit

Catalog No.: BZEK2865

Size: 96T

Range: 125 pg/ml - 2000 pg/ml

Sensitivity: 10 pg/ml

Storage and Expiration: Store at 2-8℃ for 6 months. 

Application: For quantitative detection of ADP in monkey serum, plasma, tissue homogenate or cell culture supernatant.

Introduction

Adiponectin (also referred to as GBP-28, apM1, AdipoQ and Acrp30) is a protein hormone and adipokine, which is involved in regulating glucose levels and fatty acid breakdown.[5][6] In humans, it is encoded by the ADIPOQ gene and is produced primarily in adipose tissue, but also in muscle and even in the brain. Adiponectin is a 244-amino-acid-long polypeptide (protein). It has four distinct regions: The first is a short signal sequence that targets the hormone for secretion outside the cell; next is a short region that varies between species; the third is a 65-amino acid region with similarity to collagenous proteins; the last is a globular domain. Overall, this protein shows similarity to the complement 1Q factors (C1Q), but when the three-dimensional structure of the globular region was determined, a striking similarity to TNFα was observed, despite unrelated protein sequences.

Principle of the Assay  

This kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. The purified anti- ADP antibody was pre-coated onto 96-well plates. And the HRP conjugated anti- ADP antibody was used as detection antibodies. The standards, test samples and HRP conjugated detection antibody were added to the wells subsequently, mixed and incubated, then, unbound conjugates were washed away with wash buffer. TMB substrates (A & B) 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 ADP amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of ADP can be calculated.

Kit components

1. One 96-well plate pre-coated with anti- monkey ADP antibody

2. Standard: 0.5 ml (4000 pg/ml)

3. Standard diluent buffer: 1.5 ml

4. Wash buffer (30×): 20 ml. Dilution: 1:30

5. Sample diluent buffer: 6 ml

6. HRP conjugated anti- monkey ADP antibody (RTU): 6ml

7. Stop solution: 6 ml

8. TMB substrate A: 6 ml

9. TMB substrate B: 6 ml

10. Plate sealer: 2

11. Hermetic bag: 1

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. Absorbent filter papers
8. Plastic or glass container with volume of above 1L