Measured by its ability to hydrolyze the substrate 4-Nitrocatechol Sulfate (PNCS). The specific activity is >0.25 pmol/min/µg, as measured under the described conditions.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived mouse Sulfamidase/SGSH protein Arg23-Leu502, with a C-terminal 10-His tag
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.
Applications/Dilutions
Dilutions
Enzyme Activity
Theoretical MW
56 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
60 kDa, reducing conditions
Publications
Read Publications using 2969-SU in the following applications:
Substrate: 4-Nitrocatechol Sulfate (PNCS), 100 mM stock in deionized water
Sodium Hydroxide (NaOH), 2 M stock in deionized water
Clear 96-well Plate
Plate Reader
Dilute rmSGSH to 40 µg/mL in Assay Buffer.
Dilute Substrate to 20 mM in Assay Buffer.
Combine 150 µL of 40 µg/mL rmSGSH and 150 µL of 20 mM Substrate. Include a Substrate Blank containing 150 µL of Assay Buffer and 150 µL of 20 mM Substrate.
Incubate at 37 °C for 72 hours.
Stop reactions and Substrate Blank by adding 300 µL of 0.2 M NaOH.
Load in a plate 200 µL of each reaction and Substrate Blank.
Read at 510 nm (absorbance) in endpoint mode.
Calculate specific activity:
Specific Activity (pmol/min/µg) =
Adjusted Abs* (OD) x Conversion Factor** (pmol/OD)
Incubation time (min) x amount of enzyme (µg)
*Adjusted for Substrate Blank
**Derived using calibration standard P-Nitrocatechol (PNC).
Per Well:
rmSGSH: 2 µg
Substrate: 5 mM
Notes
This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.
Alternate Names for Recombinant Mouse Sulfamidase/SGSH Protein, CF
heparan sulfate sulfatase
HSS
HSSN-sulphoglucosamine sulphohydrolase
MPS3A
MPS3Asulphamidase
N-sulfoglucosamine sulfohydrolase
SFMD
SFMDEC 3.10.1.1
SGSH
Sulfamidase
Sulfoglucosamine sulfamidase
Sulphamidase
Background
Also known as N-sulfoglucosamine sulfohydrolase and heparan N-sulfatase, Sulfamidase/SGSH is an important member of the sulfatase family involved in the degradation of heparan sulfate (1). The SGSH deficiency results in mucopolysaccharidosis type IIIA (MPS IIIA, Sanfilippo A syndrome), an autosomal recessive lysosomal storage disease characterized by neurological dysfunction but relatively mild somatic manifestations (2). The deduced amino acid sequence of mouse SGSH consists of a signal peptide (residues 1-22) and a mature chain (residues 23-502) (1). Recombinant Mouse Sulfamidase/SGSH corresponds to the mature chain and has sulfatase activity as described in the Activity section.
Diez-Roux, G. and Ballabio, A. (2005) Annu. Rev. Genomics Hum. Genet. 6:355.
Gliddon, B.L. et al. (2004) Mol. Genet. Metab. 83:239.
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