Human Iduronate 2 sulfatase/IDS HEK293 Overexpression Lysate: 製品の情報
This Human Iduronate 2 sulfatase/IDS overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Iduronate 2 sulfatase/IDS protein (Cat: 10337-H08H) from the overexpression lysate was verified.
A DNA sequence encoding human IDS precursor (NP_000193.1) (Met 1-Pro 550) was expressed with a C-terminal polyhistidine tag.
The secreted recombinant human IDS with the propeptide consists of 536 amino acids after removal of the signal peptide and has a predicted molecular mass of 61 kDa. In SDS-PAGE under reducing conditions, it migrates as several bands with apparent molecular mass between 85-95 KDa due to glycosylation.
Human Iduronate 2 sulfatase/IDS HEK293 Overexpression Lysate: 用法
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).
安定性 & 保存条件
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 Iduronate 2 sulfatase/IDS HEK293 Overexpression Lysate: 別名
Human IDS Overexpression Lysate; Human MPS2 Overexpression Lysate; Human SIDS Overexpression Lysate
Iduronate 2 sulfatase/IDS 背景情報
Iduronate 2-Sulfatase, also known as IDS, is a member of the highly conserved sulfatase family of enzymes that catalyze the hydrolysis of O- and N-sulfate esters from a variety of substrates. The human Iduronate 2-Sulfatase/IDS consists of a signal peptide, a pro peptide and a mature chain that may be further processed into two chains. Among the identified 18 human sulfatases, Iduronate 2-Sulfatase/IDS is required for the lysosomal degradation of the glycosaminoglycans (GAG), heparan sulfate and dermatan sulfate. Multiple mutations in this X-chromosome localized gene result in Iduronate 2-Sulfatase/IDS enzymatic deficiency, and lead to the sex-linked Mucopolysaccharidosis Type II (MPS II ), also known as Hunter Syndrome characterized by the lysosomal accumulation of the GAG and their excretion in urine. MPS II has a wide spectrum of clinical manifestations ranging from mild to severe due to the level of Iduronate 2-Sulfatase/IDS enzyme. Retroviral-mediated Iduronate 2-Sulfatase/IDS gene transfer into lymphoid cells would be a promising gene therapeutic strategy.
Wilson P.J., et al.,(1990), Hunter syndrome: isolation of an iduronate-2-sulfatase cDNA clone and analysis of patient DNA. Proc. Natl. Acad. Sci. U.S.A. 87:8531-8535.
Wilson P.J., et al., (1993), Sequence of the human iduronate 2-sulfatase (IDS) gene.Genomics 17:773-775.
Timms K.M., et al.,(1995), 130 kb of DNA sequence reveals two new genes and a regional duplication distal to the human iduronate-2-sulfate sulfatase locus.Genome Res. 5:71-78.