This Human CD79 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD79 protein (Cat: CT081-H2508H) from the overexpression lysate was verified.
A DNA sequence encoding the human CD79A (NP_001774.1) (Met1-Arg143) was fused with a flag tag at the C-terminus, constructed the plasmid 1; A DNA sequence encoding human CD79B (NP_000617.1) (Met1-Asp159) was fused with a polyhistidine tag at the C-terminus, constructed the plasmid 2. The two plasmids were co-expressed and the human CD79A & CD79B heterodimer was purified.
The recombinant human CD79A & human CD79B heterodimer comprises 324 (152+172) amino acids and has a calculated molecular mass of 37.5 (17.4+ 20.1) kDa.
Human CD79 HEK293 Overexpression Lysate: 使用指南
Cell lysate was prepared by homogenization 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.
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.
3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
稳定性 & 储存条件
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 CD79 HEK293 Overexpression Lysate: 别称
Human IGA Overexpression Lysate; Human MB-1 Overexpression Lysate