Human OLFM4 HEK293 Overexpression Lysate

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Human OLFM4 HEK293 Overexpression Lysate: Product Information

Product Description
This Human OLFM4 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of OLFM4 protein (Cat: 11639-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human OLFM4 (NP_006409.3) (Met 1-Gln 510) was fused with a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant human OLFM4 consists of 501 amino acids and predictes a molecular mass of 56.6 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rh OLFM4 is approximately 65-70 kDa due to different glycosylation.

Human OLFM4 HEK293 Overexpression Lysate: Usage Guide

Preparation Method
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.
Lysis Buffer
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.
Recommend Usage
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.
Sample Buffer
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.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human OLFM4 HEK293 Overexpression Lysate: Alternative Names

Human bA209J19.1 Overexpression Lysate; Human GC1 Overexpression Lysate; Human GW112 Overexpression Lysate; Human hGC-1 Overexpression Lysate; Human hOLfD Overexpression Lysate; Human OlfD Overexpression Lysate; Human OLM4 Overexpression Lysate; Human UNQ362 Overexpression Lysate

OLFM4 Background Information

Olfactomedin-4, also known as G-CSF-stimulated clone 1 protein, Antiapoptotic protein GW112, and OLFM4, is a secreted protein which contains one olfactomedin-like domain. The OLFM4 gene was recently reported to inhibit various apoptotic pathways and promote proliferation of cancer cells, suggesting that OLFM4 might serve as a diagnostic marker for human cancers. Thus, OLFM4 mRNA might be a useful tool to support the diagnosis of cancer, irrespective of the clinical stages. It is overexpressed in a number of human tumor types, especially in those of the digestive system. GW112 is associated with GRIM-19, a protein known to be involved in regulating cellular apoptosis. Functionally, GW112 could significantly attenuate the ability of GRIM19 to mediate retinoic acid-IFN-beta-mediated cellular apoptosis and apoptosis-related gene expression. In addition, GW112 demonstrated strong antiapoptotic effects in tumor cells treated with other stress exposures such as hydrogen peroxide. Finally, forced overexpression of GW112 in murine prostate tumor cells led to more rapid tumor formation in a syngeneic host. OLFM4 is an important regulator of cell death that plays important roles in tumor cell survival and tumor growth. As a candidate gene for cancer-specific expression. The serum olfactomedin 4 (OLFM4) is a useful marker for Gastric cancer (GC) and its measurement alone or in combination with Reg IV has utility in the early detection of GC. GW112 has an antiapoptotic property against the cytotoxic agents-induced apoptosis. It suggested that GW112 could be an important mediator in NF kappaB-dependent tumorigenesis of digestive tract tissues.
Full Name
olfactomedin 4
References
  • Zhang X, et al. (2004) GW112, a novel antiapoptotic protein that promotes tumor growth. Cancer Res. 64(7): 2474-81.
  • Koshida S, et al. (2007) Specific overexpression of OLFM4(GW112/HGC-1) mRNA in colon, breast and lung cancer tissues detected using quantitative analysis. Cancer Sci. 98(3): 315-20.
  • Oue N, et al. (2009) Serum olfactomedin 4 (GW112, hGC-1) in combination with Reg IV is a highly sensitive biomarker for gastric cancer patients. Int J Cancer. 125(10): 2383-92.
  • Kim KK, et al. (2010) Up regulation of GW112 Gene by NF kappaB promotes an antiapoptotic property in gastric cancer cells. Mol Carcinog. 49(3): 259-70.
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