Human VEGF-C ELISA Kit

Catalog No.: BEK1230

Size: 96T

Range: 62.5 pg/ml - 4000 pg/ml

Sensitivity < 3 pg/ml

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

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

Introduction

Vascular endothelial growth factor C (VEGF-C) is a member of the platelet-derived growth factor/vascular endothelial growth factor (PDGF/VEGF) family. It is active in angiogenesis, lymphangiogenesis and endothelial cell growth and survival, and can also affect the permeability of blood vessels. This secreted protein undergoes a complex proteolytic maturation, generating multiple processed forms that bind and activate VEGFR-3 receptors. Karkkainen et al. (2004) found that VEGFC and VEGF, unlike VEGFB and VEGFD, are essential for embryonic survival and lymphangiogenesis.

Principle of the Assay  

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

Kit components

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