Anti-Claudin 3 antibody [EPR19971] (ab214487)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19971] to Claudin 3
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
- Reacts with: Human
Overview
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Product name
Anti-Claudin 3 antibody [EPR19971]
See all Claudin 3 primary antibodies -
Description
Rabbit monoclonal [EPR19971] to Claudin 3 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanICC/IF HumanIHC-P HumanIP HumanWB Human -
Immunogen
This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: SW480, MCF7 and Caco-2 whole cell lysates; human breast cancer, pancreas, ovary cancer and fetal kidney lysates. IHC-P: Human pancreas, prostate and breast cancer tissues. ICC/IF: MCF7 cells. Flow Cyt: MCF7 cells. IP: MCF7 whole cell lysate.
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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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19971 -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-Claudin 3 antibody [EPR19971] (ab214487) at 1/1000 dilution
Lane 1 : SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate
Lane 2 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lane 3 : Caco-2 (Human colorectal adenocarcinoma cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 23 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondBlocking/Dilution buffer: 5% NFDM/TBST.
Doublet bands observed correspond to full length and processed forms of the claudin-3 protein. This is consistent with what has been described in the literature (PMID: 26170992).
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All lanes : Anti-Claudin 3 antibody [EPR19971] (ab214487) at 1/1000 dilution
Lane 1 : Human breast cancer lysate
Lane 2 : Human pancreas lysate
Lane 3 : Human ovary cancer lysate
Lane 4 : Human fetal kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/5000 dilution
Predicted band size: 23 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1/2: 1 minute; Lane 3: 3 seconds; Lane 4: 3 minutes.
Doublet bands observed correspond to full length and processed forms of the claudin-3 protein. This is consistent with what has been described in the literature (PMID: 26170992).
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Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling Claudin 3 with ab214487 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membrane staining on the pancreatic acinar cells of human pancreas is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded human prostate tissue labeling Claudin 3 with ab214487 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membrane staining on the epithelium of human benign prostatic hyperplasia is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling Claudin 3 with ab214487 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membrane staining on the tumor cells of human breast cancer is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 100% methanol-fixed MCF7 (Human breast adenocarcinoma cell line) cells labeling Claudin 3 with ab214487 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membrane staining on MCF7 cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) ab195889) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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Flow cytometric analysis of 4% paraformaldehyde-fixed MCF7 (Human breast adenocarcinoma cell line) cells labeling Claudin 3 with ab214487 at 1/50 dilution (red) compared with Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
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Claudin 3 was immunoprecipitated from 0.35 mg of MCF7 (Human breast adenocarcinoma cell line) whole cell lysate with ab214487 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab214487 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution
Lane 1: MCF7 whole cell lysate 10µg (Input).
Lane 2: ab214487 IP in MCF7 whole cell lysate.
Lane 3:Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab214487 in MCF7 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds.
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