Human HAI-1 HEK293 Overexpression Lysate: Product Information
This Human HAI-1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of HAI-1 protein (Cat: 11742-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human SPINT1 isoform 2 (O43278-2) (Met 1-Val 433) was fused with a polyhistidine tag at the C-terminus.
The recombinant human SPINT1 consists of 409 amino acids and predictes a molecular mass of 45.8 kDa. In SDS-PAGE under reducing conditions, rh SPINT1 migrates as an approximately 55 kDa band due to glycosylation.
Human HAI-1 HEK293 Overexpression Lysate: Usage Guide
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.
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