Recombinant Human Sulfamidase/SGSH Protein, CF Summary
Product Specifications
Arg23-Leu502 with a C-terminal 6-His tag
Analysis
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
8380-SU
| Formulation | Supplied as a 0.2 μm filtered solution in Tris and NaCl. |
| Shipping | The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Assay Procedure
- Assay Buffer: 50 mM MES, pH 5.5
- Recombinant Human Sulfamidase/SGSH (rhSGSH) (Catalog # 8380-SU)
- Substrate: 4-Nitrocatechol Sulfate (PNCS), 100 mM stock in deionized water
- Sodium Hydroxide (NaOH), 2 M stock in deionized water
- Clear 96-well Plate (Catalog # DY990)
- Plate Reader
- Dilute rhSGSH to 40 µg/mL in Assay Buffer.
- Dilute PNCS to 20 mM in Assay Buffer.
- Combine 150 µL of 40 µg/mL of rhSGSH and 150 µL of 20 mM of PNCS. Include a Substrate Blank containing 150 µL of Assay Buffer and 150 µL of 20 mM of PNCS.
- Incubate at 37 °C for 72 hours.
- Stop reactions and Substrate Blank by adding 300 µL of 0.2 M NaOH.
- Load into a plate 200 µL from each stopped reaction and Substrate Blank.
- Read plate 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 4-Nitrocatechol (PNC).
Scientific Data
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Recombinant Human Sulfamidase/SGSH Protein, CF (Catalog # 8380-SU) hydrolyzes N-linked sulfate group from the non-reducing terminal glucosaminide residues of heparan sulfate.
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2 μg/lane of Recombinant Human Sulfamidase/SGSH Protein (Catalog # 8380-SU) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 55-63 kDa.
Reconstitution Calculator
Background: Sulfamidase/SGSH
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 (HS) (1). Different from the HS specific endosulfatases that remove sulfate from internal GlcNAc residues (2), SGSH removes sulfate group from the non-reducing end glucosamine residues on HS. 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 (3). Human SGSH shows 88.6% sequence identity with that of mouse sequence.
- DiezRoux, G. and Ballabio, A. et al. (2005) Annu. Rev. Genomics Hum. Genet. 6:355.
- Morimoto-Tomita, M. et al. (2002) J. Biol. Chem. 277:49175.
- Blanch, L. et al. (1997) Hum. Mol. Genet. 6:787.
Citation for Recombinant Human Sulfamidase/SGSH Protein, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
1 Citation: Showing 1 - 1
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An immune deficient mouse model for mucopolysaccharidosis IIIA (Sanfilippo syndrome)
Authors: Pollock, K;Noritake, S;Imai, DM;Pastenkos, G;Olson, M;Cary, W;Yang, S;Fierro, FA;White, J;Graham, J;Dahlenburg, H;Johe, K;Nolta, JA;
Scientific reports
Species: N/A
Sample Types: Recombinant Protein
Applications: Bioassay
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