Human GLIPR1 HEK293 Overexpression Lysate: Product Information
This Human GLIPR1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of GLIPR1 protein (Cat: 10873-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human GLIPR1 (NP_006842.2) extracellular domain (Met 1-Arg 232) was expressed, with a C-terminal polyhistidine tag.
The secreted recombinant human GLIPR1 comprises 222 amino acids and has a predicted molecular mass of 25.7 kDa. rhGLIPR1 migrates as an approximately 27 kDa band in SDS-PAGE under reducing conditions.
Human GLIPR1 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 GLIPR1 HEK293 Overexpression Lysate: Alternative Names
Human CRISP7 Overexpression Lysate; Human GLIPR Overexpression Lysate; Human RTVP1 Overexpression Lysate
GLIPR1 Background Information
Glioma pathogenesis-related protein 1, also known as Protein RTVP-1, GLIPR1 and GLIPR, is a single-pass membrane protein which belongs to theCRISP family. GLIPR1 / RTVP-1 was expressed in high levels in glioblastomas, whereas its expression in low-grade astrocytomas and normal brains was very low. Transfection of glioma cells with small interfering RNAs targeting GLIPR1 / RTVP-1 decreased cell proliferation in all the cell lines examined and induced cell apoptosis in some of them. Overexpression of GLIPR1 / RTVP-1 increased astrocyte and glioma cell proliferation and the anchorage-independent growth of the cells. In addition, overexpression of GLIPR1 / RTVP-1 rendered glioma cells more resistant to the apoptotic effect of tumor necrosis factor-related apoptosis-inducing ligand and serum deprivation. GLIPR1 / RTVP-1 regulated the invasion of glioma cells was evident by their enhanced migration through Matrigel and by their increased invasion in a spheroid confrontation assay. The increased invasive potential of the GLIPR1 / RTVP-1 overexpressors was also shown by the increased activity of matrix metalloproteinase 2 in these cells. The expression of GLIPR1 / RTVP-1 is correlated with the degree of malignancy of astrocytic tumors and that GLIPR1 / RTVP-1 is involved in the regulation of the growth, survival, and invasion of glioma cells. GLIPR1 / RTVP-1 is a potential therapeutic target in gliomas.
GLI pathogenesis-related 1
Murphy E.V., et al., 1995, Gene. 159:131-5.
Rich T., et al., 1996, Gene. 180:125-30.
Rosenzweig,T. et al., 2006, Cancer Res. 66 (8):4139-48.
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