Human VEGF-D ELISA Kit

Catalog No.: BEK1231

Size: 96T

Range: 10 pg/ml-300 pg/ml

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

Application: For quantitative detection of VEGF-D in human serum, plasma, urine, cell culture supernatant or tissue samples.

Introduction

C-fos induced growth factor (FIGF) (or vascular endothelial growth factor D, VEGF-D) 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. VEGF-D has been established to be over-expressed in both tumor tissues and patients’ serum samples in several types of human cancer. In addition, VEGF-D expression has been implicated with increased incidence of regional lymph node metastasis. In experimental mice study, genetically modified tumor cell that was forced to produce VEGF-D protein have been established to boost up regional lymph nodes metastases.

Principle of the Assay  

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

Kit components

1. One 96-well plate pre-coated with anti-human VEGF-D antibody

2. Standard: 0.5ml (360pg /mL)

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-human VEGF-D 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