Rat C-Peptide, C-P ELISA Kit

Catalog No.: BYEK2527

Size: 96T

Range: 15 ng/L – 180 ng/L

Sensitivity: 1.5 ng/L

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

Application: For quantitative detection of C-P in rat serum, plasma, or cell culture supernatant.

Introduction

Connecting peptide, or C-peptide, is a short 31-amino-acid protein that connects insulin's A-chain to its B-chain in the proinsulin molecule. C-peptide has been shown to bind to the surface of a number of cell types such as neuronal, endothelial, fibroblast and renal tubular, at nanomolar concentrations to a receptor that is likely G-protein-coupled. It has been found to exert beneficial effects in many tissues affected by diabetic microvascular complications, including the kidneys. C-peptide at physiological concentrations effectively reduces diabetes-induced glomerular hyperfiltration via constriction of the afferent arteriole, dilation of the efferent arteriole, and inhibition of tubular reabsorption in experimental models of type 1 diabetes.

Principle of the Assay  

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

Kit components

1. One 96-well plate pre-coated with anti- rat C-P antibody

2. Standard: 0.5ml (270 ng/L)

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- rat C-P antibody (RTU): 6ml

7. Stop solution: 6 ml

8. TMB substrate A:  6ml

9. TMB substrate B:  6ml

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