Human S100A4 HEK293 Overexpression Lysate 产品信息
Product Description
This Human S100A4 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of S100A4 protein (Cat: 10185-H01H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the human S100A4 (NP_002952.1) (Met 1-Lys 101) was expressed with the fused Fc region of human IgG1 at the N-terminus.
Molecule Mass
The recombinant human Fc/S100A4 is a disulfide-linked homodimeric protein. The reduced monomer consists of 338 amino acids and has a predicted molecular mass of 38.4 kDa. As a result of glycosylation, the apparent molecular mass of rhFc/S100A4 monomer is approximately 40 kDa in SDS-PAGE under reducing conditions.
Human S100A4 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 S100A4 HEK293 Overexpression Lysate Alternative Names
Human 18A2 Overexpression Lysate;Human 42A Overexpression Lysate;Human CAPL Overexpression Lysate;Human FSP1 Overexpression Lysate;Human MTS1 Overexpression Lysate;Human P9KA Overexpression Lysate;Human PEL98 Overexpression Lysate
S100A4 Background Information
S1A4, also known as metastasis-associated protein Mtsl, belongs to the family of small calcium-binding S1 proteins containing two EF-hand calcium-binding motifs. In humans at least 2 S1 family members that are distributed tissue specifically have been identified, and are involved in a number of cellular processes as transducers of calcium signal. S1A4 is a symmetric homodimer, and undergoes a relatively large conformational change upon the typical EF-hand binding calcium, which is necessary for S1A4 to interact with its protein targets and generate biological effects. It can bind the already known targets p53, F-actin, liprin β, myosin heavy chain II, and prevent their phosphorylation and multimerization. It has been demonstrated that S1A4 is directly involved in tumor metastasis including cell motility, invasion, apoptosis, angiogenesis and differentiation, and appears to be a metastasis factor and a molecular marker for clinical prognosis. Multiple alternatively spliced variants encoding the same protein have been identified.
Full Name
S100 calcium binding protein A4
References
Ambartsumian N. et al., 1995, Gene. 159: 125-30. Marenholz I. et al., 2004, Biochem Biophys Res Commun. 322: 1111-22. Helfman DM. et al., 2005, Br J Cancer. 92: 1955-8. Saleem M. et al., 2006, Proc Natl Acad Sci. 103: 14825-30. Boye K. et al., 2008, Int J Cancer. 123: 1301-10. Garrett SC. et al., 2006, J Biol Chem. 281: 677-80. Kriajevska M. et al., 2002, J Biol Chem. 277: 5299-335.
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