Product Features

Product Name           Human TIMP-2 EasyTest™ ELISA Kit Assay Type                   Solid Phase Sandwich ELISA Format                           96-well strip plate Assay Process             One wash step, two incubations Assay Length              1.5 hours Sample Volume         Serum , Plasma, 50µL Detection Method    Colorimetric Sensitivity                   3pg/mL Assay Range                65.5-16000pg/mL Reactivity                     Human Cross-Reactivity       No cross-reactivity observed with available related molecules Gene Names               TIMP2; CSC-21K; DDC8 Gene ID                         7077 Swissprot ID               P16035

Recovery

Linearity

Standard Curve: Human TIMP-2 EasyTest™ ELISA Kit

Kit Components

• Pre-Coated 96-well Strip Microplate • Wash Buffer • Stop Solution • Assay Diluent • Standards • Bioaim human TIMP-2 mix • TMB One-Step Substrate

Protocol Outline

1. Prepare all reagents, samples and standards as instructed in the manual. 2. Add 50 µl of standard or sample to each well. 3. Add 50 µl of  Bioaim mix. 4. Incubate 1 h at RT. 5. Wash and add 100 µl of TMB One-Step Substrate Reagent to each well. 6. Incubate 15 min at RT. 7. Add 50 µl of Stop Solution to each well. 8. Read at 450 nm/570 nm.

Background

TIMPs-1 through -4 regulate the activity of zinc metalloproteases known as MMPs, ADAMs and ADAMTSs. Structurally, TIMPs contain two domains. The N-terminal domain binds to the active site of mature metalloproteases via a 1:1 non-covalent interaction, blocking access of substrates to the catalytic site. In addition, The C-terminal domain of TIMP-1 and TIMP-2 binds to the hemopexin- like domain of pro-MMP-9 and pro-MMP-2, respectively. The latter binding is essential for the cell surface activation of MMP-2 by MMP-14. TIMP-2 functions as both an MMP inhibitor and an activator. TIMPs inhibit all active MMPs, but different TIMPs inhibit different MMPs better than other TIMPs. For example, TIMP-1 inhibits MMP-7, MMP-9, MMP-1 and MMP-3 better than TIMP-2, and TIMP-2 inhibits MMP-2 more effectively than other TIMPs.

Storage

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Regulatory Status      

Human TIMP-2 EasyTest™ ELISA Kit is for Research Use Only.