Human EpCAM HEK293 Overexpression Lysate

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Human EpCAM HEK293 Overexpression Lysate: Product Information

Product Description
This Human EpCAM overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of EpCAM protein (Cat: 10694-H02H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the extracellular domain (Met 1-Lys265) of human EpCAM (NP_002345.1) was expressed with the fused Fc region of human IgG1 at the C-terminus.
Molecule Mass
The recombinant human EpCAM/Fc chimera is a disulfide-linked homodimeric protein after removal of the signal peptide. The monomer consists of 480 amino acids and has a calculated molecular mass of 54 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rh EpCAM/Fc is approximately 65-70 kDa due to glycosylation.

Human EpCAM 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 EpCAM HEK293 Overexpression Lysate: Alternative Names

Human DIAR5 Overexpression Lysate; Human EGP-2 Overexpression Lysate; Human EGP314 Overexpression Lysate; Human EGP40 Overexpression Lysate; Human ESA Overexpression Lysate; Human HNPCC8 Overexpression Lysate; Human KS1/4 Overexpression Lysate; Human KSA Overexpression Lysate; Human M4S1 Overexpression Lysate; Human MIC18 Overexpression Lysate; Human MK-1 Overexpression Lysate; Human TACSTD1 Overexpression Lysate; Human TROP-1 Overexpression Lysate; Human TROP1 Overexpression Lysate

EpCAM Background Information

Epithelial Cell Adhesion Molecule (EpCAM), also known as GA733-2 antigen, is a type â…transmembrane glycoprotein composed of an extracellular domain with two EGF-Like repeats and a cystenin-rich region, a transmembrane domain and a cytoplasmic domain. It modulates cell adhesion and proliferation. Its overexpression has been detected in many epithelial tumours and has been associated with high stage, high grade and a worse survival in some tumour types. EpCAM has been shown to function as a calcium-independent homophilic cell adhesion molecule that does not exhibit any obvious relationship to the four known cell adhesion molecule superfamilies. However, recent insights have revealed that EpCAM participates in not only cell adhesion, but also in proliferation, migration and differentiation of cells. In addition, recent study revealed that EpCAM is the Wnt-beta-catenin signaling target gene and may be used to facilitate prognosis. It has oncogenic potential and is activated by release of its intracellular domain, which can signal into the cell nucleus by engagement of elements of the wnt pathway.
Full Name
epithelial cell adhesion molecule
References
  • Brunner A, et al. (2008) EpCAM is predominantly expressed in high grade and advanced stage urothelial carcinoma of the bladder. J Clin Pathol. 61(3):307-10.
  • Trzpis M, et al. (2008) EpCAM in morphogenesis. Front Biosci. 13: 5050-5.
  • Munz M, et al. (2009) The emerging role of EpCAM in cancer and stem cell signaling. Cancer Res. 69(14): 5627-9.
  • Carpenter G, et al. (2009) EpCAM: another surface-to-nucleus missile. Cancer Cell. 15(3): 165-6.
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