Human TIMP-2 ELISA Kit

Catalog No.: BEK1210

Size: 96T

Range: 156 pg/ml-10,000 pg/ml

Sensitivity < 2 pg/ml

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

Application: For quantitative detection of TIMP-2 in human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

TIMP metallopeptidase inhibitor 2, also known as TIMP-2, is a 21-kD protein, which is secreted by human melanoma cells and binds to type IV collagenase proenzyme secreted by the same cells. The TIMP2 gene is a member of the TIMP gene family, It is encoded by 5 exons spanning 83 kb of genomic DNA. The proteins encoded by this gene family are natural inhibitors of the matrix metalloproteinases, a group of peptidases involved in degradation of the extracellular matrix. It has an amino acid sequence with many similarities to human tissue inhibitor of metalloproteinase (TIMP1). TIMP2 is a tissue inhibitor of metalloproteinases, TIMP2 induced a decrease in total protein tyrosine phosphatase (PTP) activity associated with beta-1 integrin subunits as well as dissociation of the phosphatase SHP1 from beta-1.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-TIMP-2 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-TIMP-2 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 TIMP-2 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of TIMP-2 can be calculated.

Kit components

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