Recombinant Human Active Heparanase/HPSE Protein, CF

Catalog # Availability Size / Price Qty
7570-GH-005
R&D Systems Recombinant Proteins and Enzymes
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Recombinant Human Active Heparanase/HPSE Protein, CF Summary

Learn more about Fluorescent Glycan Labeling and Detection

Product Specifications

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.
Activity
Measured by its ability to release biotinylated heparan sulfate from Recombinant Human Syndecan‑4 (Catalog # 2918-SD). 20 ng of Recombinant Human Active Heparanase/HPSE digestion will result in >50% of OD reduction compared with the Negative Control, as measured under the described conditions.
Source
Chinese Hamster Ovary cell line, CHO-derived human Heparanase/HPSE protein
Gln36-Ile543 with N-terminal 6-His tag
The linker peptide from Ser110-Gln157 is removed by protease treatment
Accession #
N-terminal Sequence
Analysis
His & K158
Predicted Molecular Mass
43 kDa & 9 kDa
SDS-PAGE
50-60 kDa & 7-9 kDa, reducing conditions

Product Datasheets

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7570-GH

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.

7570-GH

Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and E64.
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 Sodium Acetate, pH 5.0
  • Recombinant Human HPSE (rhHPSE) (7570-GH)
  • HPSE Substrate (Catalog # ES020)
  • Human Syndecan-4 DuoSet Kit (Catalog # DY2918)
  • 96-well Clear Plate (Catalog # DY990)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent

And the following materials that are routinely used in ELISA:

  •  Coating Buffer (Catalog # DY006)
  • Wash Buffer (25X) (Catalog # WA126)
  • Reagent Diluent (10X) (Catalog # DY995)
  • Substrate Reagent Pack (Catalog # DY999)
  • Stop Solution (Catalog # DY994)
  1. Prepare an ELISA plate by following the DuoSet kit protocol.
  2. Dilution factor determination:
    a. Dilute the HPSE Substrate stock 100-fold in Reagent Diluent (this will be the first dilution point).
    b. Further prepare a 2-fold serial dilution series of the above diluted HPSE Substrate with Reagent Diluent for 6 points.
    c. Load 100 µL of each point onto the prepared ELISA plate in duplicate. Load 100 µL of Reagent Diluent to 2 separate  wells for blank control.
    d. Cover the plate and incubate at room temperature for 2 hours.
    e. Follow the DuoSet Assay Procedure from step 4 to step 9 to complete the assay.
    f. Determine the dilution factor (n) that achieves an OD between 1.8-3.0.
  3. rhHPSE Activity Detection:
    a. Dilute the HPSE Substrate stock by n/10-fold in Assay Buffer, diute rhHPSE to 2 μg/mL in assay buffer.
    b. Combine 10 µL of rhHPSE with 10 µL of the diluted HPSE Substrate in a vial. For negative control, combine 10 µL of Assay Buffer and 10 µL of the diluted HPSE Substrate in a vial.
    c. Incubate reactions and negative control at 37 °C for 2 hours.
    d. After incubation, heat all reactions and negative control at 95 °C for 2 minutes to inactivate rhHPSE.
    e. Add 220 µL of Reagent Diluent to each reaction and negative control. Mix well.
    f. Load 100 µL of each sample onto the prepared ELISA plate in duplicate.
    g. Cover the plate and incubate for 2 hours at room temperature.
    h. Follow the DuoSet Assay Procedure form step 4 to step 9 to complete the assay.
  4. Calculate % OD reduction compared with the negative control: 

 [1-(OD of rhHPSE sample/OD of negative control)] x 100 = % OD reduction

Per Reaction:

rhHPSE: 20 ng

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: Heparanase/HPSE

Heparanase (HPSE) selectively cleaves heparan sulfate at specific sites on heparan sulfate proteoglycans (HSPGs) (1, 2, 3, 4). The enzyme is synthesized as an inactive 65 kDa proenzyme that is secreted via the Golgi apparatus and associates with the cell membrane through interaction with HSPGs (5). It is then endocytosed and transferred to lysosomes (6) where cathepsin L activates it by removing an internal inhibitory peptide, forming a heterodimer composed of an 8 kDa and a 50 kDa subunit (7, 8). Under certain stimuli, the active enzyme is transferred back to the cell surface, where it participates in extracellular matrix degradation and remodeling (9). HPSE facilitates cell migration associated with metastasis, wound healing and inflammation (10). An increase in its activity is associated with an increase in VEGF activity, which further enhances angiogenesis (11). HPSE also enhances shedding of syndecans and increases endothelial invasion and angiogenesis in myelomas (12). It acts as a procoagulant by increasing the generation of activation factor X in the presence of tissue factor and activation factor VII (13). In addition, it increases cell adhesion to the extracellular matrix (ECM), independent of its enzymatic activity (14). HPSE is highly expressed in placenta and spleen and weakly expressed in lymph node, thymus, peripheral blood leukocytes, bone marrow, endothelial cells, fetal liver and tumor tissues (15). The enzyme activity of recombinant human HPSE was assayed in an ELISA format using non-reducing end biotinylated heparan sulfate on recombinant syndecan 4 as a substrate (16).

References
  1. Vlodavsky, I. et al. (1999) Nat. Med. 5:793.
  2. Hulett, M.D. et al. (1999) Nat. Med. 5:803.
  3. Gong, F. et al. (2003) J. Biol. Chem. 278:35152.
  4. Peterson, S.B. and Liu, J. (2010) J. Biol. Chem. 285:14504.
  5. Nadav L. et al. (2002) J. Cell Sci. 115:2179.
  6. Gingis-Velitski, S. et al. (2004) J. Biol. Chem. 279:44084.
  7. Abboud-Jarrous, G. et al. (2008) J. Biol. Chem. 283:18167.
  8. Zetser, A. et al. (2004) J. Cell Sci. 117:2249.
  9. Zcharia E. et al. (2001) J. Mammary Gland Biol. Neoplasia 6:311.
  10. Fux, L. et al. (2009) Trends Biochem. Sci. 34:511.
  11. Cohen-Kaplan, V. et al. (2008) Int. J. Cancer 123:2566.
  12. Purushothaman, A. et al. (2010) Blood 115:2449.
  13. Nadir, Y. et al. (2010) Haematologica 95:1927.
  14. Goldshmidt, O. et al. (2003) FASEB J. 17:1015.
  15. Kussie, P.H. et al. (1999) Biochem. Biophys. Res. Commun. 261:183.
  16. Wu, Z.L. et al. (2017) Glycobiology 27:518.
Entrez Gene IDs
10855 (Human); 15442 (Mouse); 64537 (Rat)
Alternate Names
EC 3.2; Endo-glucoronidase; HEP; Heparanase; Heparanase-1; HPA1; HPAheparanase exon 9 and 10 deletion; HPR1; HPSE; HPSE1heparanase-1; HSE1; HSE1heparanase exon10-deletion

Citations for Recombinant Human Active Heparanase/HPSE 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.

11 Citations: Showing 1 - 10
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  1. COVID-19 generates hyaluronan fragments that directly induce endothelial barrier dysfunction
    Authors: KA Queisser, RA Mellema, EA Middleton, I Portier, BK Manne, F Denorme, EJ Beswick, MT Rondina, RA Campbell, AC Petrey
    JCI Insight, 2021;0(0):.
    Species: Human
    Sample Types: Plasma
    Applications: Bioassay
  2. Perlecan in the Natural and Cell Therapy Repair of Human Adult Articular Cartilage: Can Modifications in This Proteoglycan Be a Novel Therapeutic Approach?
    Authors: J Garcia, HS McCarthy, JH Kuiper, J Melrose, S Roberts
    Biomolecules, 2021;11(1):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  3. A Marine lambda-Oligocarrageenan Inhibits Migratory and Invasive Ability of MDA-MB-231 Human Breast Cancer Cells through Actions on Heparanase Metabolism and MMP-14/MMP-2 Axis
    Authors: R Cousin, H Groult, C Manseur, R Ferru-Clém, M Gani, R Havret, C Toucheteau, G Prunier, B Colin, F Morel, JM Piot, I Lanneluc, K Baranger, T Maugard, I Fruitier-A
    Marine Drugs, 2021;19(10):.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  4. Calcium dobesilate reduces VEGF signaling by interfering with heparan sulfate binding site and protects from vascular complications in diabetic mice
    Authors: F Njau, N Shushakova, H Schenk, VC Wulfmeyer, R Bollin, J Menne, H Haller
    PLoS ONE, 2020;15(1):e0218494.
    Species: Human
    Sample Types: Whole Cells
  5. Ischemic stroke disrupts the endothelial glycocalyx through activation of proHPSE via acrolein exposure
    Authors: K Ko, T Suzuki, R Ishikawa, N Hattori, R Ito, K Umehara, T Furihata, N Dohmae, RJ Linhardt, K Igarashi, T Toida, K Higashi
    J Biol Chem, 2020;0(0):.
    Species: Human
    Sample Types: Reference Standard
    Applications: Bioassay
  6. Heparanase-2 protects from LPS-mediated endothelial injury by inhibiting TLR4 signalling
    Authors: Y Kiyan, S Tkachuk, K Kurselis, N Shushakova, K Stahl, D Dawodu, R Kiyan, B Chichkov, H Haller
    Sci Rep, 2019;9(1):13591.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  7. Assessment of Heparanase-Mediated Angiogenesis Using Microvascular Endothelial Cells: Identification of ?-Carrageenan Derivative as a Potent Anti Angiogenic Agent
    Authors: N Poupard, P Badarou, F Fasani, H Groult, N Bridiau, F Sannier, S Bordenave-, C Kieda, JM Piot, C Grillon, I Fruitier-A, T Maugard
    Mar Drugs, 2017;15(5):.
    Applications: Bioassay
  8. 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, 2017;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Click Chemistry
  9. Production of heparin and ?-carrageenan anti-heparanase derivatives using a combination of physicochemical depolymerization and glycol splitting
    Authors: N Poupard, H Groult, J Bodin, N Bridiau, S Bordenave-, F Sannier, JM Piot, I Fruitier-A, T Maugard
    Carbohydr Polym, 2017;166(0):156-165.
    Applications: Bioassay
  10. Heparanase Activates Antithrombin through the Binding to Its Heparin Binding Site
    Authors: Nataliya Bohdan
    PLoS ONE, 2016;11(6):e0157834.
    Species: Human
    Sample Types: Whole Cells
    Applications: Western Blot
  11. Syndecan-3 and TFPI colocalize on the surface of endothelial-, smooth muscle-, and cancer cells.
    Authors: Tinholt M, Stavik B, Louch W, Carlson C, Sletten M, Ruf W, Skretting G, Sandset P, Iversen N
    PLoS ONE, 2015;10(1):e0117404.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay

FAQs

  1. What is the best practice for storage of 7570-GH at -20 oC to -70 oC?

    • 7570-GH should be stored at -20 oC to -70 oC undiluted in the vial received or undiluted as 5 µL aliquots. 7570-GH could also be diluted 5-fold in the formulation buffer (specified on the Certificate of Analysis) and stored diluted as 5 µL or larger aliquots at -20 oC to -70 oC.

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