Human CD299 HEK293 Overexpression Lysate

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Human CD299 HEK293 Overexpression Lysate: Product Information

Product Description
This Human CD299 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD299 protein (Cat: 10559-H01H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the extracellular domain (Ser 78-Glu 399) of human DC-SIGNR (NP_055072.3) was fused with the Fc region of human IgG1 at the N-terminus.
Molecule Mass
The recombinant human DC-SIGNR/Fc is a disulfide-linked homodimer after removal of the signal peptide. The reduced monomer consists of 580 amino acids and has a predicted molecular mass of 65 kDa. As a result of glycosylation, the apparent molecular mass of rhDC-SIGNR/Fc is approximately 110-140 kDa & 65-70 kDa corresponding to the homodimer and monomer respectively in SDS-PAGE under non-reduced conditions.

Human CD299 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 CD299 HEK293 Overexpression Lysate: Alternative Names

Human CD209L Overexpression Lysate; Human CD299 Overexpression Lysate; Human CLEC4M Overexpression Lysate; Human DC-SIGN2 Overexpression Lysate; Human DC-SIGNR Overexpression Lysate; Human DCSIGNR Overexpression Lysate; Human HP10347 Overexpression Lysate; Human L-SIGN Overexpression Lysate; Human LSIGN Overexpression Lysate; Human MGC129964 Overexpression Lysate; Human MGC47866 Overexpression Lysate

CD299 Background Information

C-type lectin domain family 4, member M, also known as DC-SIGNR and CLEC4M, is a type II integral membrane protein that is 77% amino acid identical to DC-SIGN, an HIV gp12-binding protein. Though the encoded gene located in the same chromosome, DC-SIGN is expressed solely on dendritic cells, while DC-SIGNR is predominantly found in liver sinusoidal endothelial cells and lymph node, as well as placental endothelium. DC-SIGNR exists as a homotetramer, and the tandem repeat domain, also called neck domain, mediates oligermerization. DC-SIGNR is ragarded as a pathogen-recognition receptor involved in peripheral immune surveillance in liver, and probably mediate the endocytosis of pathogens which are subsequently degraded in lysosomal compartments. DC-SIGNR appears to selectively recognize and bind many viral surface glycoproteins containing high mannose N-linked oligosaccharides in a calcium-dependent manner, including HIV-1 gp12, HIV-2 gp12, SIV gp12, ebolavirus glycoproteins, HCV E2, and human SARS coronavirus protein S, as well as the cellular adhesion protein ICAM3. DC-SIGNR have been thought to play an important role in establishing HIV infection by enhancing trans-infection of CD4(+)T cells in the regional lymph nodes. It may affect susceptibility to HIV infection by a mechanism that is different in females and males. DC-SIGNR can bind to hepatitis C virus (HCV), and its polymorphism might affect HCV loads supporting the concept that DC-SIGNR contributes to HCV replication efficacy.
Full Name
C-type lectin domain family 4 member M
References
  • Nattermann J, et al. (2006) The tandem-repeat polymorphism of the DC-SIGNR gene in HCV infection. J Viral Hepat. 13(1): 42-6.
  • Wichukchinda N, et al. (2007) The polymorphisms in DC-SIGNR affect susceptibility to HIV type 1 infection. AIDS Res Hum Retroviruses. 23(5): 686-92.
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