Human Calnexin HEK293 Overexpression Lysate


Human Calnexin HEK293 Overexpression Lysate: 製品の情報

This Human Calnexin overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Calnexin protein (Cat: 13929-H02H) from the overexpression lysate was verified.
HEK293 Cells
A DNA sequence encoding the human CANX (P27824) (Met1-Pro481) was expressed, fused with the Fc region of human IgG1 at the C-terminus.
The recombinant human CANX/Fc is a disulfide-linked homodimer. The reduced monomer comprises 702 amino acids and has a predicted molecular mass of 79.4 kDa. The apparent molecular mass of the protein is approximately 110 kDa in SDS-PAGE under reducing conditions.

Human Calnexin HEK293 Overexpression Lysate: 用法

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.
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.
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).
安定性 & 保存条件
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 Calnexin HEK293 Overexpression Lysate: 別名

Human CNX Overexpression Lysate; Human IP90 Overexpression Lysate; Human P90 Overexpression Lysate

Calnexin 背景情報

Calnexin is a calcium-binding protein that belongs to the calreticulin family. It interacts with newly synthesized glycoproteins in the endoplasmic reticulum. Calnexin seems to play a major role in the quality control apparatus of the ER by the retention of incorrectly folded proteins. It may act in assisting protein assembly and/or in the retention within the ER of unassembled protein subunits. Associated with partial T-cell antigen receptor complexes that escape the ER of immature thymocytes, it may function as a signaling complex regulating thymocyte maturation. Additionally it may play a role in receptor-mediated endocytosis at the synapse.
  • Rajagopalan S, et al. (1994) Retention of unassembled components of integral membrane proteins by Calnexin(CANX). Science. 263(5145):387-90.
  • Lenter M, et al. (1994) The integrin chains beta 1 and alpha 6 associate with the chaperone Calnexin(CANX) prior to integrin assembly. J Biol Chem. 269(16):12263-8.
  • Pind S, et al. (1994) Retention of unassembled components of integral membrane proteins by Calnexin(CANX). Science. 263(5145):387-90.
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