Human PSGL-1 / CD162 HEK293 Overexpression Lysate 产品信息
Product Description
This Human PSGL-1 / CD162 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of PSGL-1 / CD162 protein (Cat: 10490-H03H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the extracellular domain (Met1-Val295) of human PSGL-1 precursor (AAC50061.1) was fused with the C-terminal polyhistidine-tagged Fc region of human IgG1 at the C-terminus.
Molecule Mass
The recombinant human PSGL-1/Fc is a disulfide-linked homodimer after removal of the signal peptide and the prppeptide. The reduced monomer consists of 526 amino acids and has a predicted molecular mass of 57.1 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rh PSGL-1/Fc monomer is approximately 110-120 kDa due to glycosylation.
Human PSGL-1 / CD162 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 PSGL-1 / CD162 HEK293 Overexpression Lysate Alternative Names
Human CD162 Overexpression Lysate;Human CLA Overexpression Lysate;Human PSGL-1 Overexpression Lysate;Human PSGL1 Overexpression Lysate
PSGL-1 / CD162 Background Information
P-selectin glycoprotein ligand-1 (PSGL-1), also known as SELPLG or CD162, is the high affinitycounter-receptor for P-selectin on expressed on activated endothelial cells and platelets. PSGL-1 is a mucin-type glycoprotein, expressed on leukocytes and platelets as a homodimer of two disulfide-linked subunits of ~12 kD. As cell adhesion molecules, multiple studies have shown that PSGL-1/ P-selectin interaction is required for the normal recruitment of leukocytes during inflammatory reactions, and also participates in hemostatic responses. PSGL-1 protein requires two distinct posttranslational modifications for the Ca2+-dependent recognition by the lectin domain of P-selectin, that is tyrosine sulfation and specific O-linked glycosylation (sialic acid and fucose). PSGL-1 can also bind to other two members of the selectin family, E-selectin (endothelial) and L-selectin (leukocyte), but binds best to P-selectin.
Full Name
selectin P ligand
References
1. Sako, D. et al., 1993, Cell. 75: 1179-1186. 2. Wilkins, P. P. et al., 1995, J. Biol. Chem. 270: 22677-22680. 3. Frenette, P. S. et al., 2000, J. Exp. Med. 191: 1413-1422. 4. Vandendries, E.R .et al., 2004, Thromb. Haemost. 92: 459-466.. 5. Pouyani, T. et al., 1995, Cell. 83: 333-343.
新浪微博
腾讯微博
QQ空间
客服
