TIMP1 ELISA Kit (Dog)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Tissue Inhibitors Of Metalloproteinase 1 (TIMP1) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Tissue Inhibitors Of Metalloproteinase 1 (TIMP1) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

Metalloproteinase inhibitor 1; TIMP-1; tissue inhibitor of matrix metalloproteinase-1; tissue inhibitor of metalloproteinase 1; tissue inhibitor of metalloproteinases 1.

Metalloproteinase inhibitor that functions by forming one to one complexes with target metalloproteinases, such as collagenases, and irreversibly inactivates them by binding to their catalytic zinc cofactor. Acts on MMP1, MMP2, MMP3, MMP7, MMP8, MMP9, MMP10, MMP11, MMP12, MMP13 and MMP16. Does not act on MMP14. Also functions as a growth factor that regulates cell differentiation, migration and cell death and activates cellular signaling cascades via CD63 and ITGB1. Plays a role in integrin signaling.

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Tissue Inhibitors Of Metalloproteinase 1 (TIMP1) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Tissue Inhibitors Of Metalloproteinase 1 (TIMP1) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Tissue Inhibitors Of Metalloproteinase 1 (TIMP1), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Tissue Inhibitors Of Metalloproteinase 1 (TIMP1) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Matrix Recovery range (%) Average (%) Serum (n=5) 83-99 88 EDTA Plasma (n=5) 80-99 91 Heparin Plasma (n=5) 95-103 101 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 83-99 88 Serum (n=5) 83-99 88 EDTA Plasma (n=5) 80-99 91 EDTA Plasma (n=5) 80-99 91 Heparin Plasma (n=5) 95-103 101 Heparin Plasma (n=5) 95-103 101

Component Amount Anti-TIMP1 Microplate 96 Wells (12 x 8 Well strips) TIMP1 Lyophilized Standard 2 100X Biotinylated TIMP1 Detector Antibody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL Component Amount Component Amount Anti-TIMP1 Microplate 96 Wells (12 x 8 Well strips) Anti-TIMP1 Microplate 96 Wells (12 x 8 Well strips) TIMP1 Lyophilized Standard 2 TIMP1 Lyophilized Standard 2 100X Biotinylated TIMP1 Detector Antibody 1 x 120 uL 100X Biotinylated TIMP1 Detector Antibody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL

This assay has high sensitivity and excellent specificity for detection of Tissue Inhibitors Of Metalloproteinase 1 (TIMP1) . No significant cross-reactivity or interference between Tissue Inhibitors Of Metalloproteinase 1 (TIMP1) and analogues was observed.