Human TSP2 ELISA Kit

Catalog No.: BEK1221

Size: 96T

Range: 625 pg/ml - 40,000 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 TSP2 in human serum, plasma, body fluids, tissue lysates or cell culture supernates.

Introduction

Thrombospondin-2 is a protein that in humans is encoded by the THBS2 gene. This gene maps to 6q27, and it was transcribed in fibroblasts, smooth muscle cells, and an osteosarcoma cell line, at somewhat lower levels than that of thrombospondin I. Thrombospondin-2 belongs to the thrombospondin family, It is a disulfide-linked homotrimeric glycoprotein that mediates cell-to-cell and cell-to-matrix interactions. This protein has been shown to function as a potent inhibitor of tumor growth and angiogenesis.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-TSP2 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-TSP2 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 TSP2 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of TSP2 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-Human TSP2 antibody
2. Lyophilized Human TSP2 standards: 2 tubes (40ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Human TSP2 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