This Human VSIG4 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of VSIG4 protein (Cat: 12163-H02H) from the overexpression lysate was verified.
A DNA sequence encoding the human VSIG4 (NP_009199.1) (Met1-Pro283) was expressed with the Fc region of human IgG1 at the C-terminus.
The recombinant human VSIG4/Fc is a disulfide-linked homodimer. The reduced monomer comprises 505 amino acids and has a predicted molecular mass of 56.2 kDa. The apparent molecular mass of the protein is approximately 57 kDa in SDS-PAGE under reducing conditions.
Human VSIG4 HEK293 Overexpression Lysate: 使用指南
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).
稳定性 & 储存条件
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 VSIG4 HEK293 Overexpression Lysate: 别称
Human CRIg Overexpression Lysate; Human Z39IG Overexpression Lysate
VSIG4 (V-set and immunoglobulin domain containing 4), also known as complement receptor of the immunoglobulin superfamily (CRIg) and Z39Ig, is a type I transmembrane glycoprotein. It is a B7 family-related protein and an Ig superfamily member. In contrast to the B7 family members which contain two IgG domains, VSIG4 contains one complete V-type I g domain and a truncated C-type I g domain. VSIG4 is exclusively expressed on tissue resident macrophages and binds to multimers of C3b and iC3b that are covalently attached to particle surfaces. No VSIG4 expression appears to be present in T and B cells. VSIG4 functions as a negative regulator of T cell activation, and may be involved in the maintenance of peripheral T cell tolerance, and is also identified as a potent suppressor of established inflammation. Mouse VSIG4 is synthesized as a 28 amino acid precursor that contains a signal sequence, a V-type I g domain (aa 36-115), one potential N-linked glycosylation site, and a single transmembrane domain. The V-type I g domain of mouse VSIG4 shares 86% and 8% aa sequence identity with the V-type I g domains of rat and human VSIG4, respectively.
V-set and immunoglobulin domain containing 4
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