Recombinant Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)
![Western blot - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)](https://www.abcam.com/ps/products/288/ab288438/Images/ab288438-480201-anti-heparanase-1-antibody-epr2530358-western-blot-lysate8.jpg "Western blot - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR25303-58] to Heparanase 1
- Suitable for: IP, WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Heparanase 1 antibody [EPR25303-58]
See all Heparanase 1 primary antibodies -
Description
Rabbit monoclonal [EPR25303-58] to Heparanase 1 -
Host species
Rabbit -
Tested applications
Suitable for: IP, WB, IHC-Pmore details
Unsuitable for: Flow Cyt (Intra) or ICC -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, Capan-1, B16-F10, Human spleen, Mouse spleen and Rat spleen lysates. IHC-P: Human spleen, Human cervical carcinoma, Mouse spleen, Mouse pancreatic carcinoma and Rat spleen. IP: Capan-1 and B16-F10 cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR25303-58 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab288438 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| IP |
1/30.
|
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| WB |
1/1000. Predicted molecular weight: 61 kDa.
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|
| IHC-P |
1/200.
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| Notes |
|---|
|
IP
1/30. |
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WB
1/1000. Predicted molecular weight: 61 kDa. |
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IHC-P
1/200. |
Target
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Function
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. -
Tissue specificity
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. -
Sequence similarities
Belongs to the glycosyl hydrolase 79 family. -
Post-translational
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 localization
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. - Information by UniProt
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Database links
- Entrez Gene: 10855 Human
- Entrez Gene: 15442 Mouse
- Entrez Gene: 64537 Rat
- Omim: 604724 Human
- SwissProt: Q9Y251 Human
- SwissProt: Q6YGZ1 Mouse
- SwissProt: Q71RP1 Rat
- Unigene: 44227 Human
see all -
Alternative names
- Endo glucoronidase antibody
- Endo-glucoronidase antibody
- HEP antibody
see all
Images
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Western blot - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)Anti-Heparanase 1 antibody [EPR25303-58] (ab288438) at 1/1000 dilution + Rat spleen tissue lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution
Predicted band size: 61 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 3 minutes
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![Western blot - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)](https://www.abcam.com/ps/products/288/ab288438/Images/ab288438-480200-anti-heparanase-1-antibody-epr2530358-western-blot-lysate7.jpg)
Western blot - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)All lanes : Anti-Heparanase 1 antibody [EPR25303-58] (ab288438) at 1/1000 dilution
Lane 1 : Mouse spleen tissue lysate
Lane 2 : Mouse pancreas tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution
Predicted band size: 61 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST
Low expression: mouse pancreas (PMID: 11406531)
Exposure time: 3 minutes
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![Western blot - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)](https://www.abcam.com/ps/products/288/ab288438/Images/ab288438-480199-anti-heparanase-1-antibody-epr2530358-western-blot-lysate6.jpg)
Western blot - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)Anti-Heparanase 1 antibody [EPR25303-58] (ab288438) at 1/1000 dilution + Human spleen tissue lysate at 20 µg
Secondary
VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution
Predicted band size: 61 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:10405343, 10395326)
Exposure time: 3 minutes
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![Western blot - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)](https://www.abcam.com/ps/products/288/ab288438/Images/ab288438-480198-anti-heparanase-1-antibody-epr2530358-western-blot-pancreas2.jpg)
Western blot - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)All lanes : Anti-Heparanase 1 antibody [EPR25303-58] (ab288438) at 1/1000 dilution
Lane 1 : Untreated Capan-1 (human pancreas adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : Capan-1 whole cell lysate treated with Protein Deglycosylation MIX II
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution
Predicted band size: 61 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST
Heparanase 1 is a glycosylated protein and can be deglycosylated by Protein Deglycosylation MIX II.
Exposure time: 3 minutes
Bands: 50, 65, 42 kDa
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![Western blot - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)](https://www.abcam.com/ps/products/288/ab288438/Images/ab288438-480197-anti-heparanase-1-antibody-epr2530358-western-blot-cervix2.jpg)
Western blot - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)All lanes : Anti-Heparanase 1 antibody [EPR25303-58] (ab288438) at 1/1000 dilution
Lane 1 : Wild-type HeLa (human cervix adenocarcinoma epithelial cell) cell lysate
Lane 2 : HPSE knockout HeLa (human cervix adenocarcinoma epithelial cell) cell lysate
Lane 3 : Capan-1 (human pancreas adenocarcinoma epithelial cell) cell lysate
Lane 4 : HeLa (human cervix adenocarcinoma epithelial cell) cell lysate
Lane 5 : MCF7 (human breast adenocarcinoma epithelial cell) cell lysate
Lane 6 : B16-F10 (Mouse melanoma mixture of spindle-shaped and epithelial-like cells) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) at 1/10000 dilution
Predicted band size: 61 kDa
Observed band size: 42,50,65 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBST
Performed under reducing conditions.
Lysates at 20µg per lane.
False colour image of Western blot: Anti-Heparanase 1 antibody [EPR25303-58] (ab288438) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red.
In Western blot, ab288438 was shown to bind specifically to Heparanase 1. Two bands were observed at 50 and 65kDa in wild-type HeLa cell lysates with no signal observed at this size in Heparanase 1 knockout cell line ab265413 (knockout cell lysate ab258456). To generate this image, wild-type and Heparanase 1 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/10000 dilution.
Negative control: MCF7(PMID: 10395325). -
![Immunoprecipitation - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)](https://www.abcam.com/ps/products/288/ab288438/Images/ab288438-9-anti-heparanase-1-antibody-epr2530358-immunoprecipitation-b16f102.jpg)
Immunoprecipitation - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)Heparanase 1 was immunoprecipitated from 0.35 mg B16-F10 (Mouse skin melanoma) whole cell lysate 10 ug with ab288438 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab288438 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.Lane 1: B16-F10 (Mouse skin melanoma) whole cell lysate 10 ug
Lane 2: ab288438 IP in B16-F10 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab288438 in B16-F10 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes
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![Immunoprecipitation - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)](https://www.abcam.com/ps/products/288/ab288438/Images/ab288438-8-anti-heparanase-1-antibody-epr2530358-immunoprecipitation-capan12.jpg)
Immunoprecipitation - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)Heparanase 1 was immunoprecipitated from 0.35 mg Capan-1 (human pancreas adenocarcinoma epithelial cell) whole cell lysate 10 ug with ab288438 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab288438 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.Lane 1: Capan-1 (human pancreas adenocarcinoma epithelial cell) whole cell lysate 10 ug
Lane 2: ab288438 IP in Capan-1 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab288438 in Capan-1 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)](https://www.abcam.com/ps/products/288/ab288438/Images/ab288438-7-anti-heparanase-1-antibody-epr2530358-immunohistochemistry-ouse-pancreas.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labelling Heparanase 1 with ab288438 at 1/200 (2.07 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Negative control: no staining on mouse pancreas. The section was incubated with ab288438 at 4°C overnight. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)](https://www.abcam.com/ps/products/288/ab288438/Images/ab288438-6-anti-heparanase-1-antibody-epr2530358-immunohistochemistry-uman-thyroid.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)Immunohistochemical analysis of paraffin-embedded Human thyroid tissue labelling Heparanase 1 with ab288438 at 1/200 (2.07 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Negative control: no staining on human thyroid. The section was incubated with ab288438 at 4°C overnight. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)](https://www.abcam.com/ps/products/288/ab288438/Images/ab288438-5-anti-heparanase-1-antibody-epr2530358-immunohistochemistry-at-spleen.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labelling Heparanase 1 with ab288438 at 1/200 (2.07 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on rat spleen. The section was incubated with ab288438 at 4°C overnight. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)](https://www.abcam.com/ps/products/288/ab288438/Images/ab288438-4-anti-heparanase-1-antibody-epr2530358-immunohistochemistry-ouse-pancreatic-car.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)Immunohistochemical analysis of paraffin-embedded Mouse pancreatic carcinoma tissue labelling Heparanase 1 with ab288438 at 1/200 (2.07 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on mouse pancreatic carcinoma. The section was incubated with ab288438 at 4°C overnight. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)](https://www.abcam.com/ps/products/288/ab288438/Images/ab288438-3-anti-heparanase-1-antibody-epr2530358-immunohistochemistry-ouse-spleen.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labelling Heparanase 1 with ab288438 at 1/200 (2.07 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on mouse spleen. The section was incubated with ab288438 at 4°C overnight. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)](https://www.abcam.com/ps/products/288/ab288438/Images/ab288438-2-anti-heparanase-1-antibody-epr2530358-immunohistochemistry-uman-cervical-carci.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)Immunohistochemical analysis of paraffin-embedded Human cervical carcinoma tissue labelling Heparanase 1 with ab288438 at 1/200 (2.07 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on human cervical carcinoma. The section was incubated with ab288438 at 4°C overnight. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)](https://www.abcam.com/ps/products/288/ab288438/Images/ab288438-1-anti-heparanase-1-antibody-epr2530358-immunohistochemistry-uman-spleen.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heparanase 1 antibody [EPR25303-58] (ab288438)Immunohistochemical analysis of paraffin-embedded Human spleen tissue labelling Heparanase 1 with ab288438 at 1/200 (2.07 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on human spleen. The section was incubated with ab288438 at 4°C overnight. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)
Protocols
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.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab288438 has not yet been referenced specifically in any publications.