Description

Nori Human TIMP2 ELISA Kit Summary

Alternative names for TIMP2: TIMP metallopeptidase inhibitor 2 ( TIMP2), TIMP-2

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background: 

Tissue inhibitor of metalloproteinases 2 (TIMP2) is encoded by TIMP2 gene and belong to TIMP family.[1] TIMP are natural inhibitors of the matrix metalloproteinases (MMP), a group of peptidases involved in degradation of the extracellular matrix. In addition to an inhibitory role against metalloproteinases, TIMP2 has a unique role among TIMP family members in its ability to directly suppress the proliferation of endothelial cells. As a result, the encoded protein may be critical to the maintenance of tissue homeostasis by suppressing the proliferation of quiescent tissues in response to angiogenic factors, and by inhibiting protease activity in tissues undergoing remodelling of the extracellular matrix. TIMP2 functions as both an MMP inhibitor and an activator. TIMPs inhibit active MMPs, but different TIMPs inhibit different MMPs better than others. 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.^[2] A more recent discovery is that TIMP2 plays an important role in hippocampal function and cognitive function. It plays a critical role in the benefit conferred to old mice when given human umbilical cord blood.^[3] In melanocytic cells TIMP2 gene expression may be regulated by MITF.^[4] TIMP-2 is thought to be a metastasis suppressor. TIMP-2 is a direct target of miR-492 that modulates cervical cancer cell invasion.[5] Myopia development in mammals is associated with reduced expression of TIMP-2, which contributes to increased degradative activity in the sclera.[6] In the resected esophageal cancer an increased mRNA expression of MMP-7, MMP-10 and TIMP-1 correlated with clinicopathologic features.[7] These genes may play a role during progression of the disease. MMP-10, MMP-7, TIMP-1, TIMP-2 were overexpressed in 73%, 85%, 55% and 42% of esophageal cancer samples, respectively.

References

1. Boone TC, et al. (1990) Proc. Natl. Acad. Sci. U.S.A. 87 (7), 2800-2804 .
2. Bourboulia D, Stetler-Stevenson WG ( 2010). Seminars in Cancer Biology. 20 (3): 161–8.
3. Joseph Castellano; et al. (2017). Nature. doi:1038/nature22067.CS1 maint: Explicit use of et al. (link)
4. Hoek KS, et al. (2008). Pigment Cell Melanoma Res. 21 (6): 665–76.
5. Liu M, et al. (2018) Mol. Carcinog. 57 (1), 32-43.
6. Liu HH, et al. (2017) Invest. Ophthalmol. Vis. Sci. 58 (4), 1971-1981.
7. Juchniewicz A, et al. (2017) Acta Biochim. Pol. 64 (2), 295-299.

Product Citations