HPSE KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 17 bp deletion in exon 1.
Endo-glucoronidase, HEP, HPA, HPA 1, HPR 1, HPSE 1, HPSE_HUMAN, HSE 1, Heparanase, Heparanase 50 kDa subunit, Heparanase-1
HPSE KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 17 bp deletion in exon 1.
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
We will provide viable cells that proliferate on revival.
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.
Heparanase 1 also known as HPA1 or HPSE is an endo-β-D-glucuronidase enzyme with a molecular mass of approximately 50 kDa. It functions mechanically by cleaving heparan sulfate (HS) chains which are long sugar chains found on the extracellular matrix and cell surfaces. This cleavage activity remodels the matrix and influences cell behavior. Heparanase 1 is mainly expressed in the placenta lymphoid tissues and various tumor cells where it plays a role in matrix degradation and cell migration.
When heparan sulfate is cleaved by Heparanase 1 it causes the release of HS-bound growth factors and cytokines making them available to cells. This enzymatic activity facilitates cell proliferation and angiogenesis affecting processes important for tissue repair and cancer progression. Heparanase 1 does not require a complex for its function but operates as a monomeric enzyme. Its activity is investigated using assays like heparanase activity assays or ELISA for measuring the enzyme's activity in different biological contexts including mouse models.
The activity of Heparanase 1 intersects significantly with the regulation of angiogenesis and cell signaling pathways such as the VEGF and FGF pathways. These pathways are important for new blood vessel formation making them significant in both normal development and cancer. Heparanase 1 interacts with proteins such as VEGF which is important for blood vessel growth and integrins which are involved in cell-matrix interactions and signaling.
Heparanase 1 is associated with tumor metastasis and inflammation. Its ability to degrade the extracellular matrix enables cancer cell invasion and metastasis. The enzyme also plays a role in inflammatory diseases where it modulates the immune response. Through these disease pathways it relates to other proteins such as matrix metalloproteinases (MMPs) which also participate in matrix remodeling during tumor progression and inflammation.
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Lanes 1-4: Merged signal (red and green). Green - Anti-Heparanase 1 antibody [EPR22365-230] ab254254 observed at 52,65 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-Heparanase 1 antibody [EPR22365-230] ab254254 Anti-Heparanase 1 antibody [EPR22365-230] 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 Human HPSE (Heparanase 1) knockout HeLa cell lysate ab258456) was used. Wild-type and Heparanase 1 knockout samples were subjected to SDS-PAGE. Anti-Heparanase 1 antibody [EPR22365-230] ab254254 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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.
All lanes: Western blot - Anti-Heparanase 1 antibody [EPR22365-230] (Anti-Heparanase 1 antibody [EPR22365-230] ab254254) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: HPSE knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human HPSE (Heparanase 1) knockout HeLa cell line (ab265413)
Lane 3: Capan-1 cell lysate at 20 µg
Lane 4: Human pancreas tissue lysate at 20 µg
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 61 kDa
Observed band size: 52 kDa, 65 kDa
Homozygous: 17 bp deletion in exon 1.
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