This Human S100A1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of S100A1 protein (Cat: 10179-H01H) from the overexpression lysate was verified.
A DNA sequence encoding the human S100A1 (NP_006262.1) (Gly 2-Ser94) was expressed with the N-terminal fused Fc region of human IgG1.
The recombinant human Fc/S100A1 is a disulfide-linked homodimeric protein. The reduced monomer consists of 330 amino acids and has a predicted molecular mass of 37.1 kDa. As a result of glycosylation, the apparent molecular mass of rhFc/S100A1 monomer is approximately 40 kDa in SDS-PAGE under reducing conditions.
Human S100A1 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 S100A1 HEK293 Overexpression Lysate: 别称
Human S100 Overexpression Lysate; Human S100-alpha Overexpression Lysate; Human S100A Overexpression Lysate
S100A1 is a Ca2+ binding protein of the EF-hand type that belongs to the S100 protein family. S100 proteins consisting of at least 19 members exist as dimers in the cytoplasm and/or nucleus of a wide range of cells, and are involved in the regulation of a number of cellular processes such as cell-cycle progression and cell differentiation. This protein has been shown to function in the processes including stimulation of Ca2+-induced Ca2+ release, inhibition of microtubule assembly, and inhibition of PKC-mediated phosphorylation. Phosphoglucomutase is a target protein whose activity is antagonistically regulated by S100A1, and recently, S100A1 is also identified as a potent molecular chaperone and a new member of the Hsp70/Hsp90 multichaperone complex. S100A1 displays a tissue-specific expression pattern with highest levels in myocardium and is considered to be an important regulator of cardiac contractility. Accordingly, reduced expression or mutations of S100A1 gene have been implicated in cardiomyopathies.