Human Semaphorin 4D HEK293 Overexpression Lysate

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Human Semaphorin 4D HEK293 Overexpression Lysate: Product Information

Product Description
This Human Semaphorin 4D overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Semaphorin 4D protein (Cat: 11825-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human SEMA4D isoform 1 (Q92854-1) extracellular domain (Met 1-Arg 734) was expressed, with a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant human SEMA4D consists of 725 amino acids and predictes a molecular mass of 80.7 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rhSEMA4D is approximately 110 kDa due to glycosylation.

Human Semaphorin 4D 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 Semaphorin 4D HEK293 Overexpression Lysate: Alternative Names

Human C9orf164 Overexpression Lysate; Human CD100 Overexpression Lysate; Human coll-4 Overexpression Lysate; Human COLL4 Overexpression Lysate; Human M-sema-G Overexpression Lysate; Human SEMAJ Overexpression Lysate

Semaphorin 4D Background Information

Semaphorin 4D (SEMA4D or CD1) is a member of the semaphorin family of proteins and an important mediator of the movement and differentiation of multiple cell types, including those of the immune, vascular, and nervous systems. VEGF and SEMA4D had a positive correlation with the malignant degree of ovarian cancer, and SEMA4D can serve as an independent prognostic factor. SEMA4D was the first semaphorin described to have immune functions and serves important roles in T cell priming, antibody production, and cell-to-cell adhesion. Proteolytic cleavage of SEMA4D from the cell surface gives rise to a soluble fragment of SEMA4D (sSEMA4D). Similar to the transmembranal form, sSEMA4D is thought to have immunoregulatory properties.
Full Name
semaphorin 4D
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