Human Glypican 3 HEK293 Overexpression Lysate

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Human Glypican 3 HEK293 Overexpression Lysate: Product Information

Product Description
This Human Glypican 3 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Glypican 3 protein (Cat: 10088-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human GPC3 (P51654-1) (Met 1-His 559) was expressed, with a C-terminal polyhistidine tag.
Molecule Mass
The secreted recombinant human GPC3 comprises 546 amino acids and has a predicted molecular mass of 62 kDa. GPC3 is composed of proteolytic cleaved two subunits linked by disulfide bonds, that are produced by proprotein convertases. As a result of glycosylation, apparent molecular mass of rhGPC3 is approximately 40 and 50-60 kDa in SDS-PAGE under reducing conditions, corresponding to the N-terminal and C-terminal fragment respectively.

Human Glypican 3 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 Glypican 3 HEK293 Overexpression Lysate: Alternative Names

Human DGSX Overexpression Lysate; Human Glypican 3 Overexpression Lysate; Human GTR2-2 Overexpression Lysate; Human MXR7 Overexpression Lysate; Human OCI-5 Overexpression Lysate; Human SDYS Overexpression Lysate; Human SGB Overexpression Lysate; Human SGBS Overexpression Lysate; Human SGBS1 Overexpression Lysate

Glypican 3 Background Information

Glypican-3, also known as Intestinal protein OCI-5, GPC3, and OCI5, is a member of the glypican family. It belongs to the glypican family and is highly expressed in lung, liver, and kidney. It is a heparan sulfate proteoglycan, which is overexpressed in various neoplasms such as hepatocellular carcinoma, malignant melanoma, and testicular yolk sac tumor, and plays an important role in cell growth and differentiation. GPC3 function is tissue dependent. In some tissues, GPC3 acts as a tumor suppressor gene, whereas in others, it acts as an oncofetal protein. Studies have shown that GPC3 is a reliable marker for hepatocellular carcinoma. The sensitivity and specificity exceeds both alpha-fetoprotein and hepatocyte-paraffin1. GPC3 immunohistochemistry can aid in the differentiation of testicular germ cell tumors, being expressed in all yolk sac tumors but not in seminomas. GPC3 expression has also been identified in some squamous cell carcinomas of the lung and clear cell carcinomas of the ovary. The role of GPC3 in melanomas is still controversial. Thus, Glypican-3 is currently regarded as a tumor marker and potential target for immunotherapy.
Full Name
glypican 3
References
  • Kandil DH, et al. (2009) Glypican-3: a novel diagnostic marker for hepatocellular carcinoma and more. Adv Anat Pathol. 16(2): 125-9.
  • Maeda D, et al. (2009) Glypican-3 expression in clear cell adenocarcinoma of the ovary. Mod Pathol. 22(6): 824-32.
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