Product nameHuman HPSE (Heparanase 1) knockout HeLa cell lysate
See all Heparanase 1 kits
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.
User storage instructions: After reconstitution, store the lysate at -80°C.
Parental Cell LineHeLa
Mutation descriptionKnockout achieved by using CRISPR/Cas9, 17 bp deletion in exon1.
Knockout validationSanger Sequencing, Western Blot (WB)
Reconstitution notesTo use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.
*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
Tested applicationsSuitable for: WBmore details
Storage instructionsStore at -80°C. Please refer to protocols.
Components 1 kit ab262420 - Human HPSE knockout HeLa cell lysate (Lyophilized) 1 x 100µg ab255929 - Human Wild Type HeLa cell lysate (Lyophilized) 1 x 100µg
STR AnalysisAmelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8,12 CSF1PO: 9, 10
FunctionEndoglycosidase 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.
Tissue specificityHighly expressed in placenta and spleen and weakly expressed in lymph node, thymus, peripheral blood leukocytes, bone marrow, endothelial cells, fetal liver and tumor tissues.
Sequence similaritiesBelongs to the glycosyl hydrolase 79 family.
modificationsProteolytically 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.
Cellular localizationLysosome 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.
- Information by UniProt
- Endo glucoronidase
KO cell lines
Our Abpromise guarantee covers the use of ab258456 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration. Predicted molecular weight: 61 kDa.|
Lane 1:Wild-type HeLa cell lysate (20 ug)
Lane 2:HPSE knockout HeLa cell lysate (20 ug)
Lane 3:Capan-1 cell lysate (20 ug)
Lane 4:Human pancreas tissue lysate (20 ug)
ab254254 was shown to specifically react with Heparanase 1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265413 (knockout cell lysate ab258456) was used. Wild-type and Heparanase 1 knockout samples were subjected to SDS-PAGE. ab254254 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Homozygous: 17 bp deletion in exon1
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab258456 has not yet been referenced specifically in any publications.