Human CD32B/Fcgr2b HEK293 Overexpression Lysate

製品の説明

This Human CD32B/Fcgr2b overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD32B/Fcgr2b protein (Cat: 10259-H08H) from the overexpression lysate was verified.

発現ホスト

HEK293 Cells

種

Human

タンパク質構築情報

A DNA sequence encoding the extracellular domain (Ala 46-Pro 217) of mature human CD32b (NP_001002274.1) protein was expressed with a N-terminal signal peptide and a C-terminal polyhistidine tag.

分子量

The recombinant mature human CD32b consists of 183 amino acids after proteolytic removal of the signal peptide and predicts a molecular mass of 20.8 kDa. By SDS-PAGE under reducing conditions, it migrates as an approximately 25-30 kDa protein due to glycosylation.

調製方法

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.

溶解バッファー

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.

おすすめの用法

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.

バッファー

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.

アプリケーション

Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.