Human CD32A HEK293 Overexpression Lysate

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Human CD32A HEK293 Overexpression Lysate: Product Information

Product Description
This Human CD32A overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD32A protein (Cat: 10374-H27H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the extracellular domain (Met 1-Met 210) of human CD32a (AAA35827.1) was fused with a C-terminal polyhistidine tagged AVI tag at the C-terminus. It is identical to FCGR2A131H/R in the reference.
Molecule Mass
The recombinant human CD32a consists of 213 amino acids after removal of the signal peptide and predicts a molecular mass of 23.6 kDa. It migrates as an approximately 32 kDa protein due to glycosylation by SDS-PAGE under reducing conditions.

Human CD32A 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 CD32A HEK293 Overexpression Lysate: Alternative Names

Human CD32 Overexpression Lysate; Human CD32A Overexpression Lysate; Human CDw32 Overexpression Lysate; Human Fc gamma RIIA Overexpression Lysate; Human FCG2 Overexpression Lysate; Human FcGR Overexpression Lysate; Human FCGR2 Overexpression Lysate; Human FCGR2A1 Overexpression Lysate; Human IGFR2 Overexpression Lysate

CD32A Background Information

Full Name
Fc fragment of IgG receptor IIa
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