Human S100A2 HEK293 Overexpression Lysate 产品信息
Product Description
This Human S100A2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of S100A2 protein (Cat: 10180-H01H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the human S100A2 (NP_005969.1) (Met 2-Pro98) was expressed with the fused Fc region of human IgG1 at the N-terminus.
Molecule Mass
The recombinant human Fc/S100A2 is a disulfide-linked homodimer. The reduced monomer consists of 334 amino acids and has a predicted molecular mass of 37.6 kDa. As a result of glycosylation, the apparent molecular mass of rh Fc/S100A2 monomer is approximately 40 kDa in SDS-PAGE under reducing conditions.
Human S100A2 HEK293 Overexpression Lysate Usage Guide
Preparation Method
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.
Lysis Buffer
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Recommend Usage
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.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
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.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.
Human S100A2 HEK293 Overexpression Lysate Alternative Names
Human CAN19 Overexpression Lysate;Human S100L Overexpression Lysate
S100A2 Background Information
The calcium-binding Protein S1A2 is a member of the S1 family of proteins containing 2 EF-hand calcium-binding motifs. S1 family genes are located as a cluster on chromosome 1q21, and S1 proteins consisting of at least 2 members are involved in the regulation of a number of cellular processes such as cell-cycle progression and cell differentiation. S1A2 was first detected in lung and kidney, and is mainly expressed in a subset of tissues and cells such as breast epithelia and liver. The S1A2 protein is a homodimer that undergoes a conformational change upon binding of calcium, and the active form functions in regulating cell proliferation and differentiation, gene transcription, and p53-dependent growth arrest and apoptosis. Accordingly, this protein is regarded as a putative tumor suppressor, and thus chromosomal rearrangements and reduced expression of S1A2 gene have been implicated in certain carcinomas.
Full Name
S100 calcium binding protein A2
References
Gimona, M. et al., 1997, J. Cell. Sci. 110: 611-621. Mueller, A. et al., 2005, J. Biol. Chem. 280: 29186-29193. Lapi, E. et al., 2006, Oncogene. 25: 3628-3637. Feng, G. et al., 2001, Cancer. Res. 61: 7999-8004. Gupta, S. et al., 2003, J. Clin. Oncol. 21: 106-112.