重组Anti-Cathepsin D抗体[EPR3056Y] (ab75811)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3056Y] to Cathepsin D
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-Cathepsin D抗体[EPR3056Y]
参阅全部 Cathepsin D 一抗 -
描述
兔单克隆抗体[EPR3056Y] to Cathepsin D -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), ICC/IF, WB, IHC-Pmore details -
种属反应性
与反应: Human -
免疫原
Synthetic peptide within Human Cathepsin D aa 50-150 (N terminal). The exact sequence is proprietary.
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阳性对照
- WB: A431, HepG2 and MCF7 whole cell lysate. MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate. IHC-P: Human thyroid cancer tissue. ICC/IF: MCF-7 cells. Flow Cyt (intra): MCF-7 cells.
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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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR3056Y -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Isotype control
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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ab75811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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Flow Cyt (Intra) |
1/20.
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ICC/IF |
1/50.
For unpurified use at 1/100 - 1/250. |
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WB |
1/1000. Detects a band of approximately 44 kDa (predicted molecular weight: 44 kDa).
For unpurified use at 1/1000 - 1/2000. |
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IHC-P | (1) |
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/500. |
说明 |
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Flow Cyt (Intra)
1/20. |
ICC/IF
1/50. For unpurified use at 1/100 - 1/250. |
WB
1/1000. Detects a band of approximately 44 kDa (predicted molecular weight: 44 kDa). For unpurified use at 1/1000 - 1/2000. |
IHC-P
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/500. |
靶标
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功能
Acid protease active in intracellular protein breakdown. Involved in the pathogenesis of several diseases such as breast cancer and possibly Alzheimer disease. -
组织特异性
Expressed in the aorta extrcellular space (at protein level). -
疾病相关
Ceroid lipofuscinosis, neuronal, 10 -
序列相似性
Belongs to the peptidase A1 family.
Contains 1 peptidase A1 domain. -
翻译后修饰
N- and O-glycosylated. -
细胞定位
Lysosome. Melanosome. Secreted, extracellular space. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. In aortic samples, detected as an extracellular protein loosely bound to the matrix (PubMed:20551380). - Information by UniProt
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数据库链接
- Entrez Gene: 1509 Human
- Omim: 116840 Human
- SwissProt: P07339 Human
- Unigene: 654447 Human
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别名
- CatD antibody
- CATD_HUMAN antibody
- Cathepsin D antibody
see all
图片
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Anti-Cathepsin D antibody [EPR3056Y] (ab75811) at 1/1000 dilution + MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 15 µg
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/20000 dilution
Predicted band size: 44 kDa
Observed band size: 14,44 kDa why is the actual band size different from the predicted? -
All lanes : Anti-Cathepsin D antibody [EPR3056Y] (ab75811) at 1/1000 dilution (Unpurified)
Lane 1 : Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 2 : CTSD knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 3 : Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lysates/proteins at 40 µg per lane.
Performed under reducing conditions.
Predicted band size: 44 kDa
Observed band size: 44 kDaLanes 1 - 4: Merged signal (red and green). Green - ab75811 observed at 44 kDa. Red - loading control, ab7291, observed at 55 kDa.
ab75811 was shown to specifically react with in wild-type A431 cells as signal was lost in CTSD knockout cells. Wild-type and CTSD knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF Milk. Ab75811 and ab7291 (Mouse anti Tubulin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Cathepsin D with purified ab75811 at 1/50 dilution (1.88 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Intracellular Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Cathepsin D with purified ab75811 at 1/20 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid cancer tissue sections labeling Cathepsin D with purified ab75811 at 1/100 dilution (0.94 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (2)
ab75811 被引用在 2 文献中.
- Fu X et al. Molecular mechanism of albumin in suppressing invasion and metastasis of hepatocellular carcinoma. Liver Int 42:696-709 (2022). PubMed: 34854209
- Ashraf Y et al. Immunotherapy of triple-negative breast cancer with cathepsin D-targeting antibodies. J Immunother Cancer 7:29 (2019). PubMed: 30717773