Recombinant Guinea Pig Asparaginase His-tag Protein, CF

R&D Systems | Catalog # 10238-AS

R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems E. coli-derived Recombinant Guinea Pig Asparaginase His-tag Protein (10238-AS)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

E. coli

Accession Number

XP_003463186

Applications

Enzyme Activity
View all Asparaginase Proteins and Enzymes »

Product Specifications

Source

E. coli-derived guinea pig Asparaginase protein
Ala2-Ile565
with an N-terminal Met and 6-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

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

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

62 kDa

SDS-PAGE

57 kDa, under reducing conditions

Activity

Measured by its ability to convert asparagine to aspartic acid.
The specific activity is >2700 pmol/min/μg, as measured under the described conditions.

Scientific Data Images for Recombinant Guinea Pig Asparaginase His-tag Protein, CF

Recombinant Guinea Pig Asparaginase His-tag Protein Binding Activity

Recombinant Guinea Pig Asparaginase His-tag Protein Binding Activity

Recombinant Guinea Pig ASPG His-tag (Catalog # 10238-AS) is measured by its ability to convert asparagine to aspartic acid.
Recombinant Guinea Pig Asparaginase His-tag Protein SDS-PAGE

Recombinant Guinea Pig Asparaginase His-tag Protein SDS-PAGE

2 μg/lane of Recombinant Guinea Pig Asparaginase His-tag was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing a band under reducing conditions at 57 kDa.

Formulation, Preparation, and Storage

10238-AS
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and TCEP.
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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: Asparaginase

L-Asparaginases catalyze the conversion of L-asparagine (Asn) to L-aspartic acid (Asp) and play important roles in amino acid metabolism of significance in medical, antimicrobial, and food industry applications (1). Guinea pig ASPG (gpASPG), also known as ASNase1, is a tetramer composed of a dimer of dimers. Each protomer contains a N-terminal domain with the active site and a C-terminal domain that can act as a lid when substrate is bound (2). gpASPG was identified as an L-asparaginase capable of inhibiting tumor growth (3, 4). It was observed that unlike most normal cells, in acute lymphatic leukemia (ALL) there is little or no detectable asparagine synthetase making cancer cells specifically sensitive to extracellular asparagine depletion with asparaginase treatment (5-7). Although bacterial asparaginases are currently available as therapeutics in ALL (8), treatment using bacterial origin asparaginase leads to immunogenic intolerance and clearance issues from the blood (9). Additionally, the bacterial asparaginases also show an unnecessary side L-glutaminase activity (10). For these reasons, a human-like enzyme would be preferred for therapeutic use (9, 11) but must have efficient hydrolysis of extracellular Asn with a Km value in the low micromolar range (2). Human ASPG is unable to match the required efficiency for therapeutic use (2, 12, 13), but gpASPG is homologous and has similar kinetic efficiency to the bacterial enzymes currently being used as therapeutics (2, 14). gpASPG may consequently make an appropriate mammalian-source therapeutic for replacement. There is significant focus on asparaginases given the broad beneficial applications for these enzymes.

References

  1. Vimal, A. and A. Kumar (2017) Biotechnol. Genet. Eng. Rev. 33:40.
  2. Schalk, A. M. et al. (2014) J. Biol. Chem. 289:33175.
  3. Kidd, J. G. (1953) J. Exp. Med. 98:565.
  4. Broome, J. D. (1963) J. Exp. Med. 118:99.
  5. Neuman, R. E. and T. A. McCoy (1956) Science 124:124.
  6. Haley, E. E. et al. (1961) Cancer Res. 21:532.
  7. Su, N. et al. (2008) Pediatr. Blood Cancer. 50:274.
  8. Salzer, W. L. et al. (2014) Ann. N.Y. Acad. Sci. 1329:81.
  9. Muller, H. J. and J. Boos (1998) Crit. Rev. Oncol. Hematol. 28:97.
  10. Chan, W. K. et al. (2014) Blood 123:3596.
  11. Dellinger, C. T. and T. D. Miale (1976) Cancer 38:1843.
  12. Belviso, S. et al. (2017) PLoS One. 12:e0178174.
  13. Karamitros, C. S. and M. Konrad (2014) J. Biol. Chem. 289:12962.
  14. Srikhanta, Y. N. et al. (2013) Biochem. Biophys. Res. Commun. 436:362.

Long Name

Asparaginase

Alternate Names

ASPG, C14orf76

Entrez Gene IDs

374569 (Human); 100723883 (Guinea Pig)

Gene Symbol

ASPG

UniProt

XP_003463186

Additional Asparaginase Products

Product Documents for Recombinant Guinea Pig Asparaginase His-tag Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Recombinant Guinea Pig Asparaginase His-tag Protein, CF

For research use only

Customer Reviews for Recombinant Guinea Pig Asparaginase His-tag Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant Guinea Pig Asparaginase His-tag Protein, CF and earn rewards!

Have you used Recombinant Guinea Pig Asparaginase His-tag Protein, CF?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Protocols

View specific protocols for Recombinant Guinea Pig Asparaginase His-tag Protein, CF (10238-AS):

Materials
  • Assay Buffer: 25 mM Tris, pH 7.5
  • Recombinant Guinea Pig Asparaginase His-tag (rgpASPG) (Catalog # 10238-AS)
  • Substrate: L-Asparagine (Sigma, Catalog # A0884), 100 mM stock in Assay Buffer
  • alpha -Ketoglutaric Acid (Sigma, Catalog # K2010), 1 M stock in deionized water
  • beta -Nicotinamide adenine dinucleotide, reduced disodium salt hydrate (NADH) (Sigma, Catalog #
  • N8129), 20 mM stock in 0.1 M Sodium Borate, pH 9.0
  • Glutamic-Oxalacetic Transaminase (Sigma, Catalog # G2751)
  • Malic Dehydrogenase (Sigma, Catalog # M2634)
  • 96-well Clear Plate (Catalog # DY990)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Prepare a Reaction Mixture containing 800 µM NADH, 800 µM alpha -Ketoglutaric Acid, 8 Unit/mL Glutamic-Oxalacetic Transaminase and 4 unit/mL Malic Dehydrogenase in Assay Buffer.
  2. Dilute L-Asparagine to 20 mM in Assay Buffer.
  3. Dilute rgpASPG to 8 ng/µL in Assay Buffer.
  4. Load in plate, 25 µL of 8 ng/µL rgpASPG and add 25 µL of Reaction Mixture to all wells. Include a Substrate Blank containing 25 µL Assay Buffer and 25 µL of Reaction Mixture.
  5. Start the reaction by adding 50 µL of 20 mM L-Asparagine to all wells.
  6. Read plate in kinetic mode for 5 minutes at an absorbance of 340 nm.
  7. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

Adjusted Vmax* (OD/min) x well volume (L) x 1012 pmol/mol x -1
ext. coeff** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (µg)

     *Adjusted for Substrate Blank
      **Using the extinction coefficient 6220 M-1cm-1
      ***Using the path correction 0.32 cm
     Note: the output of many spectrophotometers is in mOD Per Well:

  • rgpASPG: 0.2 µg
  • L-Asparagine: 10 mM
  • NADH: 200 µM
  • alpha -Ketoglutaric Acid: 200 µM
  • Glutamic-Oxalacetic Transaminase: 2 Units
  • Malic Dehydrogenase: 1 Unit

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes