Recombinant Human Arylsulfatase B/ARSB Protein, CF

Catalog #: 4415-SU Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
4415-SU-010

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Arylsulfatase B/ARSB Protein, CF Summary

Product Specifications

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

Activity

Measured by its ability to hydrolyze the substrate 4-Nitrocatechol Sulfate (PNCS). The specific activity is >3,000 pmol/min/µg, as measured under the described conditions.

Source

Chinese Hamster Ovary cell line, CHO-derived human Arylsulfatase B/ARSB protein
Ser37-Met533, with a C-terminal 10-His tag

Accession #

P15848

N-terminal Sequence
Analysis

Ser37

Predicted Molecular Mass

57 kDa

SDS-PAGE

80 kDa, reducing conditions

Product Datasheets

4415-SU

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.

4415-SU

Formulation	Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Shipping	The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 6 months from date of receipt, -20 to -70 °C as supplied. 3 months, -20 to -70 °C under sterile conditions after opening.

Assay Procedure

Materials

Assay Buffer: 50 mM MES, pH 6.5
Recombinant Human Arylsulfatase B/ARSB (rhARSB) (Catalog # 4415-SU)
Substrate: 4-Nitrocatechol Sulfate (4-PNCS) (Sigma, Catalog # N-7251)
NaOH (Sigma, Catalog # S-0899)
96-well Clear Plate (Costar, Catalog # 92592)
Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent

Dilute rhARSB to 1 µg/mL in Assay Buffer.
Dilute Substrate to 2 mM in Assay Buffer.
Combine 75 µL of 1 µg/mL rhARSB and 75 µL of 2 mM Substrate. Include a Substrate Blank containing 75 µL Assay Buffer and 75 µL Substrate.
Incubate at 37 °C for 1 hour.
Stop reaction by adding 150 µL of 0.2 M NaOH.
Load 200 µL of reaction into a plate.
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) (Sigma, Catalog # N15553).

Per Well:

rhARSB: 0.050 µg
Substrate: 0.5 mM

Reconstitution Calculator

Background: Arylsulfatase B/ARSB

ARSB is a lysosomal enzyme of the sulfatase family that functions as an N-acetylgalactosamine-4-sulfatase (1). The enzyme is also active against chondroitin sulfate and dermatan sulfate (2). Deficiencies in ARSB activity are responsible for the lysosomal storage disease mucopolysaccharidosis Type VI, also known as Maroteaux-Lamy Syndrome (3).

References