重组Anti-Heparanase 1抗体[EPR22365-230] (ab254254)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22365-230] to Heparanase 1
- Suitable for: WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-Heparanase 1抗体[EPR22365-230]
参阅全部 Heparanase 1 一抗 -
描述
兔单克隆抗体[EPR22365-230] to Heparanase 1 -
宿主
Rabbit -
经测试应用
适用于: WBmore details
不适用于: ICC/IF,IHC-P or IP -
种属反应性
与反应: Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HeLa and Capan-1 whole cell lysates; Recombinant human active heparinase 1 protein (aa36-109 & 158-543).
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常规说明
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.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR22365-230 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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KO cell lines
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KO cell lysates
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab254254于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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WB |
1/1000. Detects a band of approximately 50, 65 kDa (predicted molecular weight: 61 kDa).
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说明 |
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WB
1/1000. Detects a band of approximately 50, 65 kDa (predicted molecular weight: 61 kDa). |
靶标
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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. - Information by UniProt
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数据库链接
- Entrez Gene: 10855 Human
- Omim: 604724 Human
- SwissProt: Q9Y251 Human
- Unigene: 44227 Human
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别名
- Endo glucoronidase antibody
- Endo-glucoronidase antibody
- HEP antibody
see all
图片
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All lanes : Anti-Heparanase 1 antibody [EPR22365-230] (ab254254) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : HPSE knockout HeLa cell lysate
Lane 3 : Capan-1 cell lysate
Lane 4 : Human pancreas tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 61 kDa
Observed band size: 52,65 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab254254 observed at 52,65 kDa. Red - loading control ab8245 observed at 36 kDa.
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 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 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 (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.
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Anti-Heparanase 1 antibody [EPR22365-230] (ab254254) at 1/1000 dilution + Recombinant human active heparinase 1 protein (aa36-109 & 158-543, expressed in CHo cells), 20 ng
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 61 kDa
Exposure time: 6 secondsBlocking and dilution buffer: 5% NFDM/TBST.
This blot was developed using a higher sensitivity ECL substrate.
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All lanes : Anti-Heparanase 1 antibody [EPR22365-230] (ab254254) at 1/1000 dilution
Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : Capan-1 (human pancreas adenocarcinoma epithelial cell line) whole cell lysate
Lane 3 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate
Lane 4 : Human pancreas lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 61 kDa
Observed band size: 50,65 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
Exposure times.
Lane 1: 3 minutes; Lanes 2 -4: 78 seconds.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 16458254, 11406531).
Negative control: Human pancreas, MCF7 (PMID: 11406531, 10395325).
This blot was developed using a higher sensitivity ECL substrate.
实验方案
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.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (4)
ab254254 被引用在 4 文献中.
- Liu M et al. Heparanase modulates the prognosis and development of BRAF V600E-mutant colorectal cancer by regulating AKT/p27Kip1/Cyclin E2 pathway. Oncogenesis 11:58 (2022). PubMed: 36130926
- Biering SB et al. SARS-CoV-2 Spike triggers barrier dysfunction and vascular leak via integrins and TGF-β signaling. Nat Commun 13:7630 (2022). PubMed: 36494335
- Zhi S et al. Insulin-like growth factor 2 mRNA binding protein 2 regulates proliferation, migration, and angiogenesis of keratinocytes by modulating heparanase stability. Bioengineered 12:11267-11276 (2021). PubMed: 34753397
- Biering SB et al. SARS-CoV-2 Spike triggers barrier dysfunction and vascular leak via integrins and TGF-β signaling. bioRxiv N/A:N/A (2021). PubMed: 34931188