Human CLEC10A HEK293 Overexpression Lysate

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Human CLEC10A HEK293 Overexpression Lysate: Product Information

Product Description
This Human CLEC10A overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CLEC10A protein (Cat: 10821-H01H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human CLEC10A (Q8IUN9-2) (Gln61-His292) was expressed, with the fused Fc region of human IgG1 at the N-terminus.
Molecule Mass
The recombinant human CLEC10A/Fc is a disulfide-linked homodimer. The reduced monomer comprises 492 amino acids and has a predicted molecular mass of 54.6 kDa. The apparent molecular mass of the protein is approximately 58 kDa in SDS-PAGE under reducing conditions due to glycosylation.

Human CLEC10A 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 CLEC10A HEK293 Overexpression Lysate: Alternative Names

Human CD301 Overexpression Lysate; Human CLECSF13 Overexpression Lysate; Human CLECSF14 Overexpression Lysate; Human HML Overexpression Lysate; Human HML2 Overexpression Lysate; Human MGL Overexpression Lysate

CLEC10A Background Information

CLEC1A, also known as the macrophage galactose-type calcium-type lectins (MGLs; CD31) constitute a unique class of C-type lectins because of their specificity for galactose and its structural homologues. MGLs/CD31 is a type II transmembrane glycoproteins and is expressed on macrophages and related cells of myeloid origins, particularly immature dendritic cells (DCs). There are 2 homologues: MGL1 and MGL2 (CD31a and CD31b) in mice. MGL1/CD31a induces both the production and secretion of interleukin (IL)-1. MGL1/CD31a plays a protective role against colitis by effectively inducing IL-1 production by colonic lamina propria macrophages in response to invading commensal bacteria.
Full Name
C-type lectin domain family 10, member A
References
  • Saba K, et al. (2009) A C-type lectin MGL1/CD301a plays an anti-inflammatory role in murine experimental colitis. Am J Pathol. 174(1): 144-52.
  • Suzuki N, et al. (2002) Molecular cloning and expression of cDNA encoding human macrophage C-type lectin: its unique carbohydrate binding specificity for Tn antigen. J Immunol. 1996 ;156: 128-135.
  • Tsuiji M, et al. (2002) Molecular cloning and characterization of a novel mouse macrophage C-type lectin, mMGL2, which has a distinct carbohydrate specificity from mMGL1. J Biol Chem. 277: 28892-28901.
  • Denda-Nagai K, et al. (2002) Macrophage C-type lectin on bone marrow-derived immature dendritic cells is involved in the internalization of glycosylated antigens. Glycobiology. 12: 443-450.
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