Human RBP4 ELISA Kit

Catalog No.: BEK1194

Size: 96T

Range: 312 pg/ml-20,000 pg/ml

Sensitivity < 10 pg/ml

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

Application: For quantitative detection of RBP4 in human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Retinol binding protein 4, plasma, also known as RBP4, is protein that in humans is encoded by the RBP4 gene. The RBP4 gene resides just centromeric of the cluster of CYP2C genes on 10q24. It belongs to the lipocalin family and is the specific carrier for retinol (vitamin A alcohol) in the blood. RBP4 has recently been described as an adipokine that contributes to insulin resistance in the AG4KO mouse model. It is secreted by adipocytes, and can act as a signal to other cells, when there is a decrease in plasma glucose concentration.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-RBP4 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-RBP4 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 RBP4 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of RBP4 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-Human RBP4 antibody
2. Lyophilized Human RBP4 standards:    2 tubes (20 ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Human RBP4 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