Tissue inhibitor of metalloproteinases-1 (TIMP-1) has been demonstrated in numerous tissues and body fluids as one of the important regulatory factors in connective tissue metabolism. TIMP-1 is a glycoprotein (M.W. 32kD) that inhibits collagenases, gelatinases and stromelysin activities, and that is identical with collagenase inhibitor of • 1-anticollagenase. TIMP-1 may be quantitated using this sensitive immunoassay. The TIMP-1 immunoassay kit is a solid phase enzyme immunoassay (EIA) based on one-step sandwich immunoreaction. Upon addition of TIMP-1 containing specimen, the result is a TIMP-1 molecule being sandwiched between the solid phase and enzyme labeled antibodies. After removing unbound enzyme-labeled antibody, the well is then incubated with enzyme substrate and o-phenylene-diamine (OPD), resulting in the development of color. The activity of peroxidase enzyme is proportional to the amount of antigen, so that TIMP-1 concentration in specimens can be determined from a standard curve.The TIMP-1 immunoassay kit is useful for the determination of TIMP-1 levels in fresh human serum and other body fluids. Antibodies in the kit are also reactive with bovine TIMP-1. Contents of this kit are sufficient for assay of 96 samples, including standard curve. Testing of samples in duplicate or triplicate is strongly recommended. Analytical Sensitivity and Detection Limits Sensitivity: 1.2ng/ml Range of Detection: 1.2ng/ml to 49ng/ml Typical values: TIMP-1 values of: Serum: 183 +/-30ng/ml Platelet poor plasma 68 +/-13ng/ml Amniotic fluid 1780 +/-969 CSF 60 +/-13 Tear fluid 418 +/-145 Mixed saliva 316 +/-166 Urine 7 +/-8 Kit Components:1. Anti-TIMP-1 Coated Microplate–One (1) plate containing 96 coated wells. Microplate lid included. 2.Antibody-HRP Conjugate–One (1) 1.2ml bottle.3. Buffer (Lyophilized) – One (1) bottle bovine serum albumin in sodium phosphate buffer, sufficient for 20ml.4. Color Reagent-1 (Lyophilized) – Three (3) bottles o-Phenylenediamine (OPD), sufficient for 5ml.5. Color Reagent-2 – One (1) 20ml bottle containing 0.02% Hydrogen peroxide.6. Stop Solution – One (1) 15ml bottle containing 2 N Sulfuric acid.7. TIMP-1 Standard (Lyophilized)–One (1) bottle, sufficient to make 0.5ml TIMP-1.8. Buffered Detergent – One (1) 1ml bottle Tween-20 in sodium phosphate buffer.Storage and Stability: Store components at 4°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.