Human MSH2 Baculovirus-Insect Overexpression Lysate


Human MSH2 Baculovirus-Insect Overexpression Lysate: 产品信息

This Human MSH2 overexpression lysate was created in Baculovirus-Insect Cells and intented for use as a Western blot (WB) positive control. Purification of MSH2 protein (Cat: 14425-H20B) from the overexpression lysate was verified.
Baculovirus-Insect Cells
A DNA sequence encoding the human MSH2 (NP_000242.1) (Met1-Thr934) was expressed with a C-terminal polyhistidine-tagged GST tag at the N-terminus (his-GST).
The recombinant human MSH2 consists 1171 amino acids and predicts a molecular mass of 132.6 kDa.

Human MSH2 Baculovirus-Insect Overexpression Lysate: 使用指南

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.
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
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).
稳定性 & 储存条件
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 MSH2 Baculovirus-Insect Overexpression Lysate: 别称

Human COCA1 Overexpression Lysate; Human FCC1 Overexpression Lysate; Human HNPCC Overexpression Lysate; Human HNPCC1 Overexpression Lysate; Human LCFS2 Overexpression Lysate

MSH2 背景信息

MSH2 is a key DNA mismatch repair protein, which plays an important role in genomic stability. In addition to its DNA repair function, MSH2 serves as a sensor for DNA base analogs-provoked DNA replication errors and binds to various DNA damage-induced adducts to trigger cell cycle arrest or apoptosis. Loss or depletion of MSH2 from cells renders resistance to certain DNA-damaging agents. Therefore, the level of MSH2 determines the DNA damage response.MSH2 is a central component of the mismatch repair pathway that targets mismatches arising during DNA replication, homologous recombination (HR), and in response to genotoxic stresses.MSH2 rearrangements are involved in approximately 10% of hereditary non-polyposis colorectal cancer (HNPCC) families, and in most of the rearrangements, exon 1 is deleted. Loss of human MSH2 (hMSH2) protein might be involved in the multistep pathogenesis of hematological malignancies associated with genetic instability.
mutS homolog 2
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