Neurotransmitter Receptors, Transporters, and Ion Channels
CD44
Product Name
Human CD44 HEK293 Overexpression Lysate
Synonyms
Human CDW44 Overexpression Lysate; Human CSPG8 Overexpression Lysate; Human ECMR-III Overexpression Lysate; Human HCELL Overexpression Lysate; Human HUTCH-I Overexpression Lysate; Human IN Overexpression Lysate; Human LHR Overexpression Lysate; Human MC56 Overexpression Lysate; Human MDU2 Overexpression Lysate; Human MDU3 Overexpression Lysate; Human MIC4 Overexpression Lysate; Human Pgp1 Overexpression Lysate
Description
This Human CD44 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control.
Molecule Mass
The recombinant human CD44/Fc is a disulfide-linked homodimer. The reduced monomer comprises 441 amino acids and has a predicted molecular mass of 49.1 kDa. The apparent molecular mass of the protein is approximately 76 kDa in SDS-PAGE under reducing conditions.
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.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the human CD44 (P16070-1) (Met1-Pro220) was expressed with the Fc region of human IgG1 at the C-terminus.
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).