This Human Biglycan overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Biglycan protein (Cat: 10447-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human BGN (NP_001702.1) (Asp38-Lys368) was expressed with a polyhistidine tag at the C-terminus.
The recombinant human BGN consists of 342 amino acids and predicts a molecular mass of 38.7 kDa.
Human Biglycan 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 Biglycan HEK293 Overexpression Lysate: 别称
Human DSPG1 Overexpression Lysate; Human PG-S1 Overexpression Lysate; Human PGI Overexpression Lysate; Human SLRR1A Overexpression Lysate
Biglycan, also known as PG-S1 and BGN, is a a small leucine-rich repeat proteoglycan (SLRP). It can be detected in a variety of extracellular matrix tissues, including bone, cartilage and tendon. Biglycan consists of a protein core containing leucine-rich repeat regions and two glycosaminoglycan (GAG) chains consisting of either chondroitin sulfate (CS) or dermatan sulfate (DS). Non-glycanated forms of biglycan (no GAG chains) increase with age in human articular cartilage. Biglycan interacts with collagen, both via the core protein and GAG chains. Biglycan plays a role in the mineralisation of bone. Biglycan core protein binds to the growth factors BMP-4 and influences its bioactivity.
Fisher L.W., et al.,(1989), Deduced protein sequence of bone small proteoglycan I (biglycan) shows homology with proteoglycan II (decorin) and several nonconnective tissue proteins in a variety of species. J. Biol. Chem. 264:4571-4576.
Fisher L.W., et al., (1991), Human biglycan gene. Putative promoter, intron-exon junctions, and chromosomal localization.J. Biol. Chem. 266:14371-14377.
Mallon A.-M., et al.,(2000), Comparative genome sequence analysis of the Bpa/Str region in mouse and man.Genome Res. 10:758-775.