Human ARHI HEK293 Overexpression Lysate


Human ARHI HEK293 Overexpression Lysate: 产品信息

This Human ARHI overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of ARHI protein (Cat: 14239-H04H) from the overexpression lysate was verified.
HEK293 Cells
A DNA sequence encoding the human DIRAS3 (O95661) (Met1-Lys225) was expressed with the Fc region of mouse IgG1 at the N-terminus.
The recombinant human DIRAS3/mFc comprises 461 amino acids and has a predicted molecular mass of 52.1 kDa. The apparent molecular mass of the monomer is approximately 62 kDa in SDS-PAGE under reducing conditions due to glycosylation.

Human ARHI HEK293 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 ARHI HEK293 Overexpression Lysate: 别称

Human ARHI Overexpression Lysate; Human DIRAS3 Overexpression Lysate; Human NOEY2 Overexpression Lysate; Human RHOI Overexpression Lysate

ARHI 背景信息

ARHI, also known as DIRAS3, belongs to the small GTPase superfamily, Di-Ras family. ARHI gene is a novel tumor suppressor gene located on chromosome 1p31. Downregulation of ARHI expression has been detected in many types of cancer. ARHI is expressed in normal ovarian and breast epithelial cells but not in ovarian and breast cancers. As a suppressor, ARHI is not only an important factor in the pathogenesis of gastric cancer, but also a potential factor for tumor aggravation. ARHI expression in gastric cancer can be employed to indicate favorable prognosis for the disease.
DIRAS family, GTP-binding RAS-like 3
  • Pei XH. et al., 2011, Cell Biol Int. 35 (10): 1019-24.
  • Lin D. et al., 2011, J Int Med Res. 39 (5): 1870-5.
  • Wang W. et al., 2012, Oncol Rep. 27 (6): 1953-9.
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