This Human B3GALTL overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of B3GALTL protein (Cat: 13957-H07H) from the overexpression lysate was verified.
A DNA sequence encoding the human B3GALTL (JC8008) (Ser29-Phe494) was expressed with an N-terminal polyhistidine tag.
The recombinant human B3GALTL comprises 486 amino acids and has a predicted molecular mass of 55.4 kDa. The apparent molecular mass of the protein is approximately 54 kDa in SDS-PAGE under reducing conditions due to glycosylation.
Human B3GALTL 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 B3GALTL HEK293 Overexpression Lysate: 别称
Human B3GALTL Overexpression Lysate; Human B3GTL Overexpression Lysate
B3GALTL is a beta-1,3-glucosyltransferase that transfers glucose to O-linked fucosylglycans on thrombospondin type-1 repeats (TSRs) of several proteins. B3GALTL is a type II membrane protein. Defects in B3GALTL gene are a cause of Peters-plus syndrome (PPS). As an O-fucosyltransferase, B3GALTL transfers glucose toward fucose with a beta-1,3 linkage. It specifically glucosylates O-linked fucosylglycan on TSP type-1 domains of proteins, thereby contributing to elongation of O-fucosylglycan.
Heinonen TY. et al., 2003, Biochem Biophys Res Commun. 309 (1): 166-74.
Sato T. et al., 2006, Glycobiology. 16 (12): 1194-206.
Kozma K. et al., 2006, J Biol Chem. 281 (48): 36742-51.