Catalog Number: INS-26-1-0000-21 (850 μg)
BACKGROUND
Heparanase is an endo-β-D-glucuronidase, which degrades heparan
sulfate side chains of heparan sulfate proteoglycans (HSPGs) in the
extracellular matrix. Heparanase plays an important role in ECM
degradation, facilitating the migration and extravasation of tumor cells
and inflammatory leukocytes (1,2,3). Upon degradation, Heparanase
releases growth factors and cytokines that stimulate cell proliferation
and chemotaxis (4,5).
Heparanase is a heterodimer comprised of a 50 kDa subunit harboring
the active site and an 8 kDa subunit. It is produced as a latent 65 kDa
precursor and proteolytically processed to its active form (1,6).
Heparanase is highly expressed in myeloid leukocytes (i.e.
neutrophils) in platelets and in human placenta. Human Heparanase
was found to be upregulated in various types of primary tumors,
correlating in some cases with increased tumor invasiveness and
vascularity and with poor prospective survival (7,8).
SOURCE
Mab HP130 is a Protein G affinity purified monoclonal antibody
raised against the 65 kDa Heparanase precursor. It recognizes the Cterminal
region of both the latent pro-Heparanase and the active
heterodimeric enzyme.
Ig CLASS
Mouse IgG1κ
PRODUCT
Each vial contains 850 μg of Mab HP130 in 500 μl of 0.22 micron
filtered solution of 20 mM Sodium Phosphate; 150 mM NaCl; pH 7.2,
containing 0.01% Thimerosal.
APPLICATIONS
Mab HP130 was shown to be active in:
FACS and Immunofluorescence analysis (9,10)
Immunohistochemistry (IHC) (11,12,13,14,15,16, 20)
Western blot (7,11,12,17,19,21)
Immunoprecipitation (IP) (18)
SPECIFICITY
In immunoblot analysis Mab HP130 reacts with the 50 kDa subunit
and with the 65 kDa precursor of human Heparanase.
Western blot analysis: recommended dilution range 1:200 (8.5 μg/ml).
IHC: recommended dilution range: 1:20 (85 μg/ml).
The antibodies cross react with the chicken Heparanase.
PURITY
>95% on SDS-PAGE.
STORAGE
Store at 4°C. Stable for six months from the date of shipment. For
extended storage, freeze in working aliquots at -20°C. Avoid
repeated freeze-thaw cycles.
RESEARCH USE
For in vitro research use only. Not for use in diagnostic
procedures.
PATENTS
Anti-Heparanase antibodies and their uses, including Mab HP130
and its uses, are protected by US. Patents No. 6,177,545;
6,531,129, additional US patent applications and patents and
patent applications worldwide.
REFERENCES
1. I. Vlodavsky, Y. Friedmann, M. Elkin, H. Aingorn, R. Atzmon,
R. Ishai-Michaeli, M. Bitan, O. Pappo, T. Peretz, I. Michal, L.
Spector, I. Pecker. 1999. Mammalian Heparanase: gene cloning,
expression and function in tumor progression and metastasis. Nat.
Med. 5: 793–802.
2. I. Vlodavsky, Y. Friedman. 2001. Molecular properties and
involvement of Heparanase in cancer metastasis and angiogenesis. J.
Clin. Invest. 108: 341–347.
3. C.R. Parish, C. Freeman, M.D. Hulett. 2001. Heparanase: a key
enzyme involved in cell invasion. Biochem. Biophys. Acta 1471:
M99–M108.
4. I. Vlodasvsky, G. Korner, R. Ishai-Michaeli, P. Bashkin, R. Bar-
Shavit, Z. Fuks, 1990. Extracellular matrix-resident growth factors
and enzyme: Possible involvement in tumor metastasis and
angiogenesis. Cancer Metastasis Rev. 9: 203-226.
5. P. Bashkin, S. Doctrow, M. Klagsbrun, C.M. Svahn, J. Folkman,
I. Vlodavsky. 1989. Basic fibroblast growth factor binds to
subendothelial extracellular matrix and is released by heparitinase and
heparin-like molecules. Biochemistry 28: 1737-1743.
6. M.B. Fairbanks, A.M. Mildner, J.W. Leone, G.S. Cavey, W.R.
Mathews, R.F. Drong, J.L. Slightom, M.J. Bienkowski, C.W. Smith,
C.A. Bannow, R.L. Heinrikson. 1999. Processing of the human
Heparanase precursor and evidence that the active enzyme is a
heterodimer. J. Biol. Chem. 274: 29587– 29590.
7. A. Koliopanos, H. Friess, J. Klee., X. Shi, Q. Liao, I. Pecker, I.
Vlodavsky, A. Zimmermann, M.W. Buchler. 2001. Heparanase
expression in primary and metastatic pancreatic cancer. Cancer Res.
61: 4655–4659.
8. K. Gohji, H. Hirano, M. Okamoto, S. Kitazawa, M. Toyoshima,
J. Dong, Y. Katsuoka, M. Nakajima. 2001. Expression of three
extracellular matrix degradative enzymes in bladder cancer. Int. J.
Cancer 95: 295–301.
9. M. Bitan, A. Polliack, G. Zecchina, A. Nagler, Y. Friedmann, L.
Nadav, V. Deutsch, I. Pecker, A. Eldor, I. Vlodavsky, and B.Z. Katz.
2002. Heparanase expression in human leukemias is restricted to
acute myeloid leukemias. Exp. Hematol. 30: 34–41.
10. S. Benhamron, H. Nechushtan, I. Verbovetski, A. Krispin, G.
Abboud-Jarrous, E. Zcharia, E. Edovitsky, E. Nahari,T. Peretz, I
Vlodavsky, and D. Mevorach. 2006. Translocation of active
Heparanase to cell surface regulates degradation of extracellular
Monoclonal Anti-Human Heparanase 1 (HPA1) Antibody
Clone HP130
InSight Biopharmaceuticals Ltd. Rabin Science Park, P.O.B. 2128 Rehovot 76121, Israel
Tel. (972)-8-9463032 Fax. (972)-8-9461042
www.insight.co.il
InSight
matrix heparan sulfate upon transmigration of mature monocytederived
dendritic cells. J. Immunol. 176: 6417-6424.
11. L. Nadav, A. Eldor, O.Yacoby-Zeevi, E. Zamir, I. Pecker, N.
Ilan, B. Geiger, I. Vlodavsky, and B.Z. Katz. 2002. Activation,
processing and trafficking of extracellular Heparanase by primary
human fibroblasts. J. Cell Science 115, 2179-2187.
12. J.P. Li, M. L. Escobar Galvis, F. Gong, X. Zhang,, E. Zcharia, S.
Metzger, I.Vlodavsky, R. Kisilevsky, and U. Lindahl. 2005. In vivo
fragmentation of heparan sulfate by Heparanase overexpression
renders mice resistant to amyloid protein amyloidosis. Proc. Natl.
Acad. Sci. 3(102): 6473-6477.
13. O. Goldshmidt, R. Yeikilis, N. Mawasi, M. Paizi, N. Gan, N.
Ilan, O. Pappo, I. Vlodavsky and G. Spira. 2004. Heparanase
expression during normal liver development and following partial
hepatectomy. J. Pathol. 203: 594–602
14. S. Gingis-Velitski, A. Zetser, V. Kaplan, O. Ben-Zaken, E.
Cohen, F. Levy-Adam, Y. Bashenko, M. Y. Flugelman, I. Vlodavsky,
and N. Ilan. 2004. Heparanase uptake is mediated by cell membrane
heparan sulfate proteoglycans. J. Biol. Chem. 279(42): 44084-44092.
15. O. Goldshmidt, L Nadav, H. Aingorn, I. Cohen, N. Feinstein,
N. Ilan, E. Zamir, B. Geiger, I. Vlodavsky and B. Z. Katz. 2002.
Human Heparanase is localized within lysosomes in a stable form.
Exp. Cell Res. 281:50–62.
16. O. Goldshmidt, E. Zcharia, H. Aingorn, Z. Guatta-Rangini, R.
Atzmon, I. Michal, I. Pecker, E. Mitrani and I. Vlodavsky. 2001.
Secretion and expression pattern of human and chicken Heparanase
are determined by their signal peptide sequences. J. Biol. Chem. 276.
17. Y. Xiao. J. Kleeff , X. Shi, M.W. Büchler, and H. Friess. 2003.
Heparanase expression in hepatocellular carcinoma and the cirrhotic
Liver. Hepatology Res. 26:192 -198.
18. A. Zetser, F. Levy-Adam, V. Kaplan, S. Gingis-Velitski, Y.
Bashenko, S. Schubert, M. Y. Flugelman, I. Vlodavsky, and N. Ilan.
2004. Processing and activation of latent Heparanase occurs in
lysosomes . J. Cell Sci. 117: 2249-2258.
19. E. Zcharia, R. Zilka, A. Yaar, O.Yacoby-Zeevi, A. Zetser, S.
Metzger, R. Sarid, A. Naggi, B. Casu, N. Ilan, I. Vlodavsky and R.
Abramovitch. 2005. Heparanase accelerates wound angiogenesis and
wound healing in mouse and rat models. FASEB J. 19: 211-221.
20. V. Temkin, H. Aingorn, I. Puxeddu, O. Goldshmidt, E. Zcharia,
G. J. Gleich, I. Vlodavsky, and F. Levi-Schaffer. 2004. Eosinophil
major basic protein: First identified natural Heparanase-inhibiting
protein. J Allergy Clin. Immunol. 113: 703-709.
21. P. Beckhove, B. M. Helmke, Y. Ziouta, M. Bucur, W. Dorner,
C. Mogler, G. Dyckhoff, and C. Herold-Mende. 2005. Heparanase
expression at the invasion front of human head and neck cancers and
correlation with poor prognosis. Clin. Cancer Res. 11(08): 2899-
2906.
PERFORMANCE
A. Reduced LDS-PAGE of Mab HP130
Monoclonal anti-human Heparanase-1 (HPA1) antibody clone HP130
(4 μg) was separated on a 12% bis-Tris SDS-polyacrylamide gel
electrophoresis (NuPage, Invitrogen) followed by GelCode Blue®
(Pierce) staining. Sample was prepared with DTT (reducing
conditions). Arrows indicate the position of heavy and light chain
bands.
B. Immunoblot analysis using Mab HP130
Purified 65 kDa precursor Heparanase (50 ng; lane 1) and purified 50
kDa Heparanase subunit (50 ng; lane 2) were loaded onto 4-12% SDSPAGE.
The proteins were transferred to PVDF membrane and
subjected to Western blot analysis using HP130. The 65 kDa precursor
and the 50 kDa subunits are clearly detected.
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