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.
功能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.
组织特异性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.
序列相似性Belongs to the glycosyl hydrolase 79 family.
翻译后修饰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.
细胞定位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.
Predicted band size : 62 kDa Observed band size : 62 kDa
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 to the generation of linker peptides, 8 kDa and 50 kDa product (SwissProt).
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.
Hong L et al. Expression of heparanase is associated with breed-specific morphological characters of placental folded bilayer between Yorkshire and Meishan pigs. Biol Reprod90:56 (2014).
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