This Human ATP1B1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of ATP1B1 protein (Cat: 14255-H07H) from the overexpression lysate was verified.
A DNA sequence encoding the human ATP1B1 (P05026-1) (Glu63-Ser303) was expressed with an N-terminal polyhistidine tag.
The recombinant human ATP1B1 comprises 261 amino acids and has a predicted molecular mass of 30.4 kDa. The apparent molecular mass of the protein is approximately 40-47 kDa in SDS-PAGE under reducing conditions due to glycosylation.
Human ATP1B1 HEK293 Overexpression Lysate: 使用指南
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.
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.
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 ATP1B1 HEK293 Overexpression Lysate: 别称
Human ATP1B Overexpression Lysate
ATP1B1 belongs to the family of Na+/K+ and H+/K+-ATPases beta chain proteins, and to the subfamily of Na+/K+ -ATPases. ATP1B1 is a subunit of Na+/K+-ATPase. Na+/K+-ATPase is an integral membrane protein responsible for establishing and maintaining the electrochemical gradients of Na and K ions across the plasma membrane. Na+/K+-ATPase is composed of two subunits, a large catalytic subunit (alpha) and a smaller glycoprotein subunit (beta). ATP1B1 regulates, through assembly of alpha/beta heterodimers, the number of sodium pumps transported to the plasma membrane. ATP1B1 is the non-catalytic component of the active enzyme, which catalyzes the hydrolysis of ATP coupled with which catalyzes the hydrolysis of ATP coupled with the exchange of Na+ and K+ ions across the plasma membrane.
ATPase, Na+/K+ transporting, beta 1 polypeptide
Lingrel JB. et al., 1990, Prog Nucleic Acid Res Mol Biol. 38: 37-89.