Rat VEGF ELISA Kit

Catalog No.: BEK1228

Size: 96T

Range: 15.6 pg/ml-1000 pg/ml

Sensitivity < 1 pg/ml

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

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

Introduction

Vascular endothelial growth factor (VEGF) is encoded by VEGF gene, which contains 8 exons and maps to 6p21.3. It is a signal protein produced by cells that stimulates vasculogenesis and angiogenesis. Millauer et al. (1994) observed in Rat that its expression was upregulated by hypoxia and that its cell surface receptor, Flk1, is exclusively expressed in endothelial cells. Overexpression of VEGF can cause vascular disease in the retina of the eye and other parts of the body. Drugs such as bevacizumab can inhibit VEGF and control or slow those diseases. The VEGF is a candidate hormone for facilitating glucose passage across the blood-brain barrier under critical conditions.

Principle of the Assay  

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

Kit components

1. One 96-well plate pre-coated with anti-Rat VEGF antibody
2. Lyophilized Rat VEGF standards: 2 tubes (10 ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Rat VEGF 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