Arylsulfatase A/ARSA Protein, Human (HEK293, His)

Arylsulfatase A (ARSA) protein specializes in hydrolyzing cerebroside sulfate, demonstrating its key role in breaking down this particular substrate. Arylsulfatase A/ARSA Protein, Human (HEK293, His) is the recombinant human-derived Arylsulfatase A/ARSA protein, expressed by HEK293 , with C-6*His labeled tag.

Arylsulfatase A (ARSA) protein is specialized in the hydrolysis of cerebroside sulfate. This enzymatic function involves the cleavage of cerebroside sulfate, showcasing ARSA's role in the breakdown of this specific substrate.

Measured by its ability to hydrolyze the substrate 4-Nitrocatechol Sulfate (PNCS). The specific activity is >25 pmol/min/µg.

Materials
Assay buffer: 50 mM NaOAc, 0.5 M NaCl, pH 4.5
Arylsulfatase A/ARSA Protein, Human (HEK293, His) (HY-P7608A)
Substrate: 4-Nitrocatechol Sulfate (PNCS) , diluted to 100 mM with ddH₂O
Standard: 4-Nitrocatechol (PNC, HY-W066890)
Sodium Hydroxide (NaOH) , 2.0 M stored in deionized water
96-well transparent plate
Microplate reader

Procedure
1. Standard Curve:Prepare 4-nitrocatechol (PNC) standards at concentrations of 0、1×10⁸、2.5×10⁸、5×10⁸、1×10⁹、2×10⁹ pmol. Measure the OD₅₁₀ for each concentration of the standard. Plot the standard curve with the concentration of the standard (pmol) on the x-axis and OD₅₁₀ on the y-axis.
2. Dilute Human ARSA to 2, 20, 200 μg/mL in the assay buffer.
3. Dilute the substrate to 2 mM in the assay buffer.
4. Experimental Group: Mix 75 μL of 2, 20, 200 μg/mL Human ARSA with 75 μL of 2 mM substrate.
Blank Control: Combine 75 μL of assay buffer with 75 μL of substrate.
Set up three replicates for each group and add to 1.5 mL centrifμge tubes.
5. Incubate in a water bath at 37°C for 1 hour.
6. Add 150 μL of 0.2 M NaOH to terminate the reaction.
7. Transfer 200 μL to a transparent 96-well plate.
8. Read the absorbance at 510 nm in endpoint mode.
9. Calculate specific activity:

Human

HEK293

C-6*His

P15289 (R19-A507)

410  [NCBI]

Full Length of Mature Protein

RPPNIVLIFADDLGYGDLGCYGHPSSTTPNLDQLAAGGLRFTDFYVPVSLCTPSRAALLTGRLPVRMGMYPGVLVPSSRGGLPLEEVTVAEVLAARGYLTGMAGKWHLGVGPEGAFLPPHQGFHRFLGIPYSHDQGPCQNLTCFPPATPCDGGCDQGLVPIPLLANLSVEAQPPWLPGLEARYMAFAHDLMADAQRQDRPFFLYYASHHTHYPQFSGQSFAERSGRGPFGDSLMELDAAVGTLMTAIGDLGLLEETLVIFTADNGPETMRMSRGGCSGLLRCGKGTTYEGGVREPALAFWPGHIAPGVTHELASSLDLLPTLAALAGAPLPNVTLDGFDLSPLLLGTGKSPRQSLFFYPSYPDEVRGVFAVRTGKYKAHFFTQGSAHSDTTADPACHASSSLTAHEPPLLYDLSKDPGENYNLLGGVAGATPEVLQALKQLQLLKAQLDAAVTFGPSQVARGEDPALQICCHPGCTPRPACCHCPDPHA

approximately 65 kDa

• 
  ≥ 95%, as determined by reducing SDS-PAGE.

Lyophilized powder

Lyophilized from a 0.22 μm filtered solution of 20 mM Tris-HCl, 150 mM NaCl, pH 8.0.
Note: For SPR assay, please replace the buffer. Primary amine components (e.g., Tris, imidazole) can affect protein-coupled chips.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).

Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Room temperature in continental US; may vary elsewhere.