Human SEMA6A HEK293 Overexpression Lysate

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Human SEMA6A HEK293 Overexpression Lysate: Product Information

Product Description
This Human SEMA6A overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of SEMA6A protein (Cat: 11189-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human SEMA6A (NP_065847.1) extracellular domain (Met 1-Thr 649) was expressed, with a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant human SEMA6A consists of 642 amino acids and predictes a molecular mass of 72.2 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rh SEMA6A is approximately 85-95 kDa due to glycosylation.

Human SEMA6A 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 SEMA6A HEK293 Overexpression Lysate: Alternative Names

Human HT018 Overexpression Lysate; Human SEMA Overexpression Lysate; Human SEMA6A1 Overexpression Lysate; Human SEMAQ Overexpression Lysate; Human VIA Overexpression Lysate

SEMA6A Background Information

SEMA6A, the first identified class 6 semaphorin, is contributed to regulate vascular development and adult angiogenesis. SEMA6A could inhibit proliferation, migration, and invasion in different glioma cell lines. That SEMA6A may be a potential prognostic biomarker in the treatment of GBM. Among different semaphorins tested by reverse transcriptase-polymerase chain reaction in human immune cells, the expression of class 6 transmembrane semaphorin SEMA6A was restricted to dendritic cells (DCs).
Full Name
sema domain, transmembrane domain (TM), and cytoplasmic domain, (semaphorin) 6A
References
  • Klostermann A., et al., 2000, J. Biol. Chem. 275:39647-53.
  • Nagase T., et al., 2000, DNA Res. 7:65-73.
  • Schmutz J., et al., 2004, Nature 431:268-74.
  • Neufeld, G. et al., 2005, Front Biosci. 10 : 751-60.
  • Fiore,R. et al., 2005, Mol Cell Biol. 25 (6):2310-9.
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