This Mouse PLA2G2D overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of PLA2G2D protein (Cat: 5A1107-M02H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse PLA2G2D (NP_035239.1) (Met1-Cys144) was expressed with the Fc region of human IgG1 at the C-terminus.
The recombinant mouse PLA2G2D consists of 363 amino acids and predicts a molecular mass of 41 kDa.
Mouse PLA2G2D HEK293 Overexpression Lysate: 用法
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).
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.