The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/500. Predicted molecular weight: 62 kDa.
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Endoglycosidase which is a cell surface and extracellular matrix-degrading enzyme. Cleaves heparan sulfate proteoglycans (HSPGs) into heparan sulfate side chains and core proteoglycans. Also implicated in the extravasation of leukocytes and tumor cell lines. Due to its contribution to metastasis and angiogenesis, it is considered to be a potential target for anti-cancer therapies.
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.
Belongs to the glycosyl hydrolase 79 family.
Proteolytically processed. The cleavage of the 65 kDa form leads to the generation of a linker peptide, 8 kDa and 50 kDa product. The active form, the 8/50 kDa heterodimer, is resistant to degradation. Complete removal of the linker peptide appears to be a prerequisite to the complete activation of the enzyme. N-glycosylated. Glycosylation of the 50 kDa subunit appears to be essential for its solubility.
Lysosome membrane. Secreted. Secreted, internalised and transferred to late endosomes/lysosomes as a proheparanase. In lysosomes, it is processed into the active form, the heparanase. The uptake or internalisation of proheparanase is mediated by HSPGs. Heparin appears to be a competitor and retain proheparanase in the extracellular medium.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Heparanase 1 antibody (ab85543)This image is courtesy of an Abreview submitted by Jim Manavis
ab85543 staining Heparanase 1 in Human lung tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% NHS for 30 minutes at room temperature; antigen retrieval was by heat mediation with an EDTA buffer. Samples were incubated with primary antibody (1/4000 in NHS) for 8 hours. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody.
Predicted band size : 62 kDa Observed band size : 62 kDa Purified recombinant human heparanase served as a control (Lane1). Lane2: An extract from 6.5x10e3 CHO cells, transfected with human Heparanase; Lane3: extract from 8x10e4 CHO cells, transfected with mouse Heparanase, and Lane4: extract from 5x10e5 non-transfected CHO dhfr- cells, were loaded onto 4-12% NuPage gel. The proteins were transferred to a PVDF and subjected to Western Blot analysis using ab85543 at 1:1000 dilution. The 65kD precursor and the 50kD and 8kD subunits are clearly detected. Lane 1 : Purified recombinant human Heparanase served as a control
Lane 2 : Extract from 6.5x10e3 CHO cells, transfected with human Heparanase
Lane 3 : Extract from 8x10e4 CHO cells, transfected with mouse Heparanase
Lane 4 : Extract from 5x10e5 non-transfected CHO dhfr- cells
Cell were loaded onto 4-12% NuPage gel. The proteins were transferred to a PVDF and subjected to Western Blot analysis using ab85543 at 1:1000 dilution.
The cleavage of the 65 kDa form leads
Zhang Y et al. Elemene inhibits the migration and invasion of 4T1 murine breast cancer cells via heparanase. Mol Med Rep16:794-800 (2017).
Read more (PubMed: 28560389) »
An X et al. Hyperoside pre-treatment prevents glomerular basement membrane damage in diabetic nephropathy by inhibiting podocyte heparanase expression. Sci Rep7:6413 (2017).
Read more (PubMed: 28743882) »