Human TIMP2 HEK293 Overexpression Lysate: Product Information
This Human TIMP2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of TIMP2 protein (Cat: 10396-HNAH) from the overexpression lysate was verified.
A DNA sequence encoding the mature form of human TIMP2 (NP_003246.1) (Cys 27-Pro 220) was expressed and purified.
The recombinant human TIMP2 consists of 195 amino acids and has a predicted molecular mass of 22 kDa. It migrates as an approximately 20 kDa band in SDS-PAGE under reducing conditions.
Human TIMP2 HEK293 Overexpression Lysate: Usage Guide
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.
2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human TIMP2 HEK293 Overexpression Lysate: Alternative Names
Human CSC-21K Overexpression Lysate; Human DDC8 Overexpression Lysate
TIMP2 Background Information
Tissue inhibitors of metalloproteinases (TIMP) family are natural inhibitors of the matrix metalloproteinases (MMPs), the zinc enzymes involved in extracellular matrix maintenance and remodeling. The TIMP family encompasses four members (TIMP1-4), and they inhibit most MMPs by forming non-covalent binary complex. TIMP2 is a 22 kDa non N-glycosylated protein expressed by a variety of cell types, and plays a unique role among TIMP family members owing to its functions to regulate cellular responses to growth factors. Findings establish an unexpected, MMP-independent mechanism for TIMP2 inhibition of endothelial cell proliferation in vitro and reveal an important component of the antiangiogenic effect of TIMP2 in vivo. TIMP-2 thus is critical to the maintenance of tissue homeostasis and is involved in the regulation of tumor microenvironment.
TIMP metallopeptidase inhibitor 2
Stetler-Stevenson, W.G. et al., 1992, Matrix. Suppl.1: 299-306.
Stetler-Stevenson, W.G. et al., 2005, Trends. Mol. Med. 11: 97-103.
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