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Recombinant P. heparinus Heparinase III Protein, CF

Catalog #: 6145-GH
8 Citations Datasheet / COA / SDS
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6145-GH-010
Recombinant P. heparinus Heparinase III Protein Enzyme Activity
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Citations (8)
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant P. heparinus Heparinase III Protein, CF Summary

Learn more about Fluorescent Glycan Labeling and Detection

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to liberate oligosaccharides from heparin. The specific activity is >3,000 pmol/min/μg, as measured under the described conditions.
Source
E. coli-derived p. heparinus Heparinase III protein
Gln25-Pro659, with an N-terminal Met and 6-His tag
Accession #
AAB18278
N-terminal Sequence
Analysis
Met
Predicted Molecular Mass
74 kDa
SDS-PAGE
66 kDa, reducing conditions

Product Datasheets

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6145-GH

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.

6145-GH

Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
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.

Assay Procedure

Materials
  • Assay Buffer: 50 mM Tris, 2 mM CaCl2, pH 7.5
  • Recombinant P. heparinus Heparinase III (rP. heparinus Heparinase III) (Catalog # 6145-GH)
  • Substrate: Heparin (Tocris, Catalog # 2812), 20 mg/mL stock in deionized water
  • 96 well clear UV-transparent microplate (Corning, Catalog # 3635)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rP. heparinus Heparinase III to 0.5 ng/µL in Assay Buffer.
  2. Dilute Substrate to 1.5 mg/mL in Assay Buffer.
  3. Load into a plate 100 µL of the diluted rP. heparinus Heparinase III, and start the reaction by adding 200 µL of 1.5 mg/mL Substrate. Include a Substrate Blank containing 100 µL of Assay Buffer and 200 µL of 1.5 mg/mL Substrate.
  4. Read in kinetic mode for 5 minutes at an absorbance of 232 nm.
  5. Calculate specific activity:

     Specific Activity (pmoles/min/µg) =

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

     *Adjusted for Substrate Blank 
     **Using the extinction coefficient 3800 M-1cm-1 
     ***Using the path correction 0.92 cm
     Note: the output of many spectrophotometers is in mOD. Per Well:
  • rP. heparinus Heparinase III: 0.05 µg
  • Substrate: 1.0 mg/mL

Scientific Data

Enzyme Activity Recombinant P. heparinus Heparinase III Protein Enzyme Activity View Larger

Heparinase III digestion of Heparan Sulfate (200 μg) is assessed in a 5-minute kinetic assay at room temperature by monitoring absorbance at 232 nm. R&D Systems RecombinantP. heparinumHeparinase III (Catalog # 6145-GH) (orange) exhibits much greater activity than the Bacteroides Heparinase III from the competition (green).

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Heparinase III

Heparan sulfate is a sulfated glycosaminoglycan with the repeating disaccharide units of -4HexA1,4GlcNAc beta 1-. It is usually attached to the protein cores of proteoglycans found on the cell membrane and in the extracellular matrix where it binds to a variety of protein ligands and regulates a wide range of biological activities (1, 2). Heparan sulfate has a domain structure where sulfated regions are interspaced with less or non-sulfated regions (3, 4). Heparin shares the backbone structure with heparan sulfate but contains no non‑sulfated regions. Heparinases are a family of lyases that release unsaturated oligosaccharides from heparin and heparan sulfate upon digestion (5). Heparinase I recognizes highly sulfated regions and is more specific for heparin. Heparinase II digests both heparin and heparan sulfate. Heparinase III prefers less-sulfated regions and is more active on heparan sulfate (6, 7).

References
  1. MacArthur, J. M. et al. (2007) J. Clin. Invest. 117:153.
  2. Esko, J. D. and Selleck, S. B. (2002) Annu. Rev. Biochem. 71:435.
  3. Maccarana, M. et al. (1996) J. Biol. Chem. 271:17804.
  4. Linker, A. and Hovingh, P. (1975) Biochim. Biophys. Acta. 385:324.
  5. Linker, A. and Hovingh, P. (1965) J. Biol. Chem. 240:3724.
  6. Su, H. et al. (1996) Appl. Environ. Microbiol. 62:2723.
  7. Hovingh, P. and Linker, A. (1970) J. Biol. Chem. 245:6170.
Entrez Gene IDs
8254931 (P. heparinus)
Alternate Names
Heparinase III

Citations for Recombinant P. heparinus Heparinase III 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.

8 Citations: Showing 1 - 8
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  1. Shifts in receptors during submergence of an encephalitic arbovirus
    Authors: Li, W;Plante, JA;Lin, C;Basu, H;Plung, JS;Fan, X;Boeckers, JM;Oros, J;Buck, TK;Anekal, PV;Hanson, WA;Varnum, H;Wells, A;Mann, CJ;Tjang, LV;Yang, P;Reyna, RA;Mitchell, BM;Shinde, DP;Walker, JL;Choi, SY;Brusic, V;Llopis, PM;Weaver, SC;Umemori, H;Chiu, IM;Plante, KS;Abraham, J;
    Nature
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  2. Targeting Chondroitin Sulfate Reduces Invasiveness of Glioma Cells by Suppressing CD44 and Integrin beta1 Expression
    Authors: YH Chu, WC Liao, YJ Ho, CH Huang, TJ Tseng, CH Liu
    Cells, 2021-12-20;10(12):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Bioassay
  3. Systematic analysis of SARS-CoV-2 infection of an ACE2-negative human airway cell
    Authors: M Puray-Chav, KM LaPak, TP Schrank, JL Elliott, DP Bhatt, MJ Agajanian, R Jasuja, DQ Lawson, K Davis, PW Rothlauf, Z Liu, H Jo, N Lee, K Tenneti, JE Eschbach, C Shema Mugi, EM Cousins, EW Cloer, HR Vuong, LA VanBlargan, AL Bailey, P Gilchuk, JE Crowe, MS Diamond, DN Hayes, SPJ Whelan, A Horani, SL Brody, D Goldfarb, MB Major, SB Kutluay
    Cell Reports, 2021-06-23;0(0):109364.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  4. Biomimetic post-capillary venule expansions for leukocyte adhesion studies
    Authors: BL Benson, L Li, JT Myers, RD Dorand, UA Gurkan, AY Huang, RM Ransohoff
    Sci Rep, 2018-06-19;8(1):9328.
  5. Imaging specific cellular glycan structures using glycosyltransferases via click chemistry.
    Authors: Wu Z, Person A, Anderson M, Burroughs B, Tatge T, Khatri K, Zou Y, Wang L, Geders T, Zaia J, Sackstein R
    Glycobiology, 2018-02-01;0(0):.
    Species: Human, Mouse
    Sample Types: Whole Cells
    Applications: Click Chemistry
  6. Novel combinatorial screening identifies neurotrophic factors for selective classes of motor neurons
    Authors: S Schaller, D Buttigieg, A Alory, A Jacquier, M Barad, M Merchant, D Gentien, P de la Gran, G Haase
    Proc. Natl. Acad. Sci. U.S.A, 2017-03-07;114(12):E2486-E2493.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  7. Calcitriol imparts neuroprotection in vitro to midbrain dopaminergic neurons by upregulating GDNF expression.
    Authors: Orme, Rowan P, Bhangal, Manminde, Fricker, Rosemary
    PLoS ONE, 2013-04-23;8(4):e62040.
    Species: Rat
    Sample Types: Whole Cells
    Applications: Bioassay
  8. Detection of specific glycosaminoglycans and glycan epitopes by in vitro sulfation using recombinant sulfotransferases.
    Authors: Wu ZL, Prather B, Ethen CM
    Glycobiology, 2010-12-17;21(5):625-33.
    Species: Bovine, Porcine
    Sample Types: Heparin, Protein
    Applications: Enzyme Assay

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