Overview

  • 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

    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Claudin 3 aa 150 to the C-terminus. The exact sequence is proprietary.
    Database link: O15551

  • 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.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab214487 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
WB 1/1000. Detects a band of approximately 20 kDa (predicted molecular weight: 23 kDa).
IHC-P 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
Flow Cyt 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

IP 1/30.

Target

  • Function

    Plays a major role in tight junction-specific obliteration of the intercellular space, through calcium-independent cell-adhesion activity.
  • Involvement in disease

    Note=CLDN3 is located in the Williams-Beuren syndrome (WBS) critical region. WBS results from a hemizygous deletion of several genes on chromosome 7q11.23, thought to arise as a consequence of unequal crossing over between highly homologous low-copy repeat sequences flanking the deleted region.
  • Sequence similarities

    Belongs to the claudin family.
  • Cellular localization

    Cell junction > tight junction. Cell membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • C7orf1 antibody
    • Claudin-3 antibody
    • Claudin3 antibody
    • CLD3_HUMAN antibody
    • CLDN 3 antibody
    • Cldn3 antibody
    • Clostridium perfringens enterotoxin receptor 2 antibody
    • CPE R2 antibody
    • CPE receptor 2 antibody
    • CPE-R 2 antibody
    • CPE-receptor 2 antibody
    • CPETR 2 antibody
    • CPETR2 antibody
    • HRVP 1 antibody
    • HRVP1 antibody
    • Rat ventral prostate 1 like protein antibody
    • Rat ventral prostate.1 protein homolog antibody
    • RVP1 antibody
    • Ventral prostate.1 like protein antibody
    • Ventral prostate.1 protein homolog antibody
    see all

Images

  • 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 second


    Blocking/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).

  • 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).

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

References

ab214487 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Western blot
Sample
Mouse Tissue lysate - other (colon)
Gel Running Conditions
Reduced Denaturing (15% gel)
Loading amount
2 µg
Specification
colon
Blocking step
PVDF Blocking Reagent (TOYOBO) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Sep 05 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rhesus monkey Tissue sections (Lung/Respiratory Epithelium)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 9.0 EDTA
Permeabilization
No
Specification
Lung/Respiratory Epithelium
Blocking step
Casein in TBS as blocking agent for 15 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Feb 02 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Colon)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 9.0 EDTA
Permeabilization
No
Specification
Colon
Blocking step
Casein as blocking agent for 15 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Feb 02 2017

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