Human B3GNT2 HEK293 Overexpression Lysate

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Human B3GNT2 HEK293 Overexpression Lysate: Product Information

Product Description
This Human B3GNT2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of B3GNT2 protein (Cat: 11416-H01H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human B3GNT2 (Q9NY97-1) (Lys29-Cys397) was expressed,with the fused Fc region of human IgG1 at the N-terminus.
Molecule Mass
The recombinant human B3GNT2/Fc is a disulfide-linked homodimer. The reduced monomer comprises 629 amino acids and has a predicted molecular mass of 71.2 kDa. The apparent molecular mass of the protein is approximately 112-120 kDa in SDS-PAGE under reducing conditions due to glycosylation.

Human B3GNT2 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 B3GNT2 HEK293 Overexpression Lysate: Alternative Names

Human B3GN-T2 Overexpression Lysate; Human B3GNT Overexpression Lysate; Human B3GNT-2 Overexpression Lysate; Human B3GNT1 Overexpression Lysate; Human BETA3GNT Overexpression Lysate; Human BGnT-2 Overexpression Lysate; Human BGNT2 Overexpression Lysate

B3GNT2 Background Information

B3GNT2 belongs to the beta-1,3-N-acetylglucosaminyltransferase family. It is a type II transmembrane protein which prefers the substrate of lacto-N-neotetraose. Alternative splicing produced 2 isoforms of the human protein. B3GNT2 catalyzes the initiation and elongation of poly-N- acetyllactosamine chains. Enzymatic activities of some glycosyltransferases are markedly increased via complex formation with other transferases or cofactor proteins. B3GNT2 and beta3Gn-T8 can form a heterodimer in vitro and that the complex exhibits much higher enzymatic activity than either enzyme alone. It is found that up-regulation of beta3Gn-T8 in differentiated HL-6 cells may increases poly-N-acetyllactosamine chains by activating intrinsic B3GNT2.
Full Name
UDP-GlcNAc:betaGal beta-1,3-N-acetylglucosaminyltransferase 2
References
  • Australo-An, et al. (2010) Genome-wide association study of ankylosing spondylitis identifies non-MHC susceptibility loci. Nat Genet. 42(2):123-7.
  • Kim W, et al. (2011) Systematic and quantitative assessment of the ubiquitin-modified proteome. Mol Cell. 44(2):325-40.
  • Seko A, et al. (2008) Activation of beta1,3-N-acetylglucosaminyltransferase-2 (beta3Gn-T2) by beta3Gn-T8. Possible involvement of beta3Gn-T8 in increasing poly-N-acetyllactosamine chains in differentiated HL-60 cells. J Biol Chem. 283(48):33094-100.
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