Human Iduronate 2 sulfatase HEK293 Overexpression Lysate 产品信息
Product Description
This Human Iduronate 2 sulfatase overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Iduronate 2 sulfatase protein (Cat: 10337-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding human IDS precursor (NP_000193.1) (Met 1-Pro 550) was expressed with a C-terminal polyhistidine tag.
Molecule Mass
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 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 Iduronate 2 sulfatase HEK293 Overexpression Lysate Alternative Names
Human IDS Overexpression Lysate;Human MPS2 Overexpression Lysate;Human SIDS Overexpression Lysate
Iduronate 2 sulfatase Background Information
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.
Full Name
iduronate 2-sulfatase
References
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.
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