Cat No.: 31103 Other Names: CLGI, EPA, Fibroblast Collagenase Inhibitor
Introduction
Tissue inhibitor of metalloproteinases-1(TIMP-1) plays a crucial role in extracellular matrix (ECM) composition and wound healing. It has been found to be strongly upregulated in liver tissue and serum during hepatic fibrogenesis in patients with liver disease and in
animal models of hepatic fibrogenesis.
Principle of the Assay
This assay is a quantitative sandwich ELISA. The immunoplate is pre-coated with a monoclonal antibody specific for human TIMP-1. Standards and samples are pipetted into the wells and any human TIMP-1 present is bound by the immobilized antibody. After
washing away any unbound substances, a biotin labelled polyclonal antibody specific for human TIMP-1 is added to the wells. After
wash step to remove any unbound reagents, streptavidin-HRP conjugate (STP-HRP) is added. After the last wash step, an HRP
substrate solution is added and colour develops in proportion to the amount of human TIMP-1 bound initially. The assay is stopped
and the optical density of the wells determined using a microplate reader. Since the increases in absorbance are directly
proportional to the amount of captured human TIMP-1, the unknown sample concentration can be interpolated from a reference
curve included in each assay.
Assay Performance
A. Typical representation of standard curve
The following standard curve is provided for demonstration only. A standard curve should be generated for each set of sample
assay.
Human TIMP-1 (pg/mL) | Absorbance (450 nm) | Blanked Absorbance |
0 | 0.091 | 0 |
31.2 | 0.118 | 0.027 |
62.5 | 0.161 | 0.07 |
125 | 0.257 | 0.166 |
250 | 0.449 | 0.358 |
500 | 0.876 | 0.785 |
1000 | 1.544 | 1.453 |
2000 | 2.494 | 2.403 |
B. Sensitivity
The lowest level of human adiponectin that can be detected by this assay is 31.2 pg/mL.
C. Precision
Intra-assay Precision (Precision within an assay) C.V.<5.0%.
Inter-assay Precision (Precision between assays) C.V.<3.9%.