Human TIMP-1 ELISA Kit

Catalog No.: BEK1208

Size: 96T

Range: 15.6 pg/ml - 1000 pg/ml

Sensitivity < 5 pg/ml

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

Application: For quantitative detection of TIMP-1 in human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Tissue inhibitor of metalloproteinase 1 (TIMP1), also known as Erythroid Potentiating Activity (EPA), is a 28,000 MW glycoprotein that is expressed from the several tissues of organisms. This protein is a member of the TIMP family. The glycoprotein is a natural inhibitor of the matrix metalloproteinases (MMPs), a group of peptidases involved in degradation of the extracellular matrix. It plays a major role in modulating the activity of interstitial collagenase as well as a number of connective tissue metalloendoproteases. Decreased TIMP1 and increased cathepsin V/L2 levels might play a role in the matrix degradation that is a hallmark of keratoconus corneas. It is able to promote cell proliferation in a wide range of cell types, and may also have an anti-apoptotic function.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-TIMP-1 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-TIMP-1 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-1 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of TIMP-1 can be calculated.

Kit components

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