Human EG-VEGF ELISA Kit

Catalog No.: BEK1046

Size: 96T

Range: 15.6 pg/ml-1000 pg/ml

Sensitivity < 7 pg/ml

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

Application: For quantitative detection of EG-VEGF in human serum, plasma, or cell culture supernatants.

Introduction

Endocrine gland-derived vascular endothelial growth factor (EG-VEGF) is a human protein encoded by the PROKR1 gene. It induces proliferation, migration, and fenestration in capillary endothelial cells derived from endocrine glands. EG-VEGF is mitogenic and chemoattractive and able to induce fenestration. Its expression is induced by hypoxia, and there is an HIF1 binding site present on EG-VEGF. It is also able to induce angiogenesis and ovarian cyst formation.

Principle of the Assay  

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

Kit components

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