HIV-1 (group M, subtype CRF07_BC) gp140 HEK293 Cell Lysate (WB positive control)

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HIV-1 (group M, subtype CRF07_BC) gp140 HEK293 Cell Lysate (WB positive control): Product Information

Product Description
This HIV HIV gp140 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of HIV gp140 protein (Cat: 11677-V08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
HIV
Sequence Information
A DNA sequence encoding the HIV-1 envelope glycoprotein gp160 extracellular domain (Val30-Tyr677), termed as gp140, was fused with the bacteriophage T4 fibritin and a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant HIV-1 gp140 consists of 694 amino acids after removal of the signal peptide and has a predicted molecular mass of 78.2 kDa. The apparent molecular mass of the recombinant protein is approximately 119 KDa in SDS-PAGE under reducing conditions.

HIV-1 (group M, subtype CRF07_BC) gp140 HEK293 Cell Lysate (WB positive control): 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.

HIV gp140 Background Information

Full Name
CUB domain containing protein 1
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