Human VCAM1 HEK293 Overexpression Lysate: Product Information
This Human VCAM1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of VCAM1 protein (Cat: 10113-H02H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain (Met 1-Pro 697) of human VCAM1 (NP_001069.1) was expressed with the Fc region of human IgG1 at the C-terminus.
The recombinant human VCAM1/Fc is a disulfide-linked homodimeric protein. The reduced monomer consists of 911 amino acids and has a predicted molecular mass of 101 kDa. As a result of glycosylation, the rh VCAM1/Fc monomer migrates as an approximately 130-140 kDa band in SDS-PAGE under reducing conditions.
Human VCAM1 HEK293 Overexpression Lysate: Usage Guide
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.
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.
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.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human VCAM1 HEK293 Overexpression Lysate: Alternative Names
Human CD106 Overexpression Lysate; Human INCAM-100 Overexpression Lysate; Human VCAM-1 Overexpression Lysate
VCAM1 Background Information
Vascular cell adhesion molecule 1 (VCAM-1), also known as CD16, is a cell surface sialoglycoprotein belonging to the immunoglobulin superfamily. Two forms of VCAM-1 with either six or seven extracellular Ig-like domains are generated by alternative splicing, with the longer form predominant. VCAM-1 is an endothelial ligand for very late antigen-4 (VLA-4) and α4ß7 integrin expressed on leukocytes, and thus mediates leukocyte-endothelial cell adhesion and signal transduction. VCAM-1 expression is induced on endothelial cells during inflammatory bowel disease, atherosclerosis, allograft rejection, infection, and asthmatic responses. During these responses, VCAM-1 forms a scaffold for leukocyte migration. VCAM-1 also activates signals within endothelial cells resulting in the opening of an "endothelial cell gate" through which leukocytes migrate. VCAM-1 has been identified as a potential anti-inflammatory therapeutic target, the hypothesis being that reduced expression of VCAM-1 will slow the development of atherosclerosis. In addition, VCAM-1-activated signals in endothelial cells are regulated by cytokines indicating that it is important to consider both endothelial cell adhesion molecule expression and function during inflammatory processes.
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