Overview

  • Product nameAnti-Claudin 3 antibody
    See all Claudin 3 primary antibodies
  • Description
    Rabbit polyclonal to Claudin 3
  • Specificityab52231 detects endogenous levels of total Claudin 3 protein.
  • Tested applicationsSuitable for: ELISA, WB, ICC/IF, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide derived from within amino acids 181 and 220 of Human Claudin-3

  • Positive control
    • Extracts from HuvEc cells

Properties

Applications

Our Abpromise guarantee covers the use of ab52231 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
ELISA 1/40000.
WB 1/500 - 1/1000. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).
ICC/IF Use a concentration of 5 µg/ml.
IP Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • FunctionPlays a major role in tight junction-specific obliteration of the intercellular space, through calcium-independent cell-adhesion activity.
  • Involvement in diseaseNote=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 similaritiesBelongs to the claudin family.
  • Cellular localizationCell 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

Anti-Claudin 3 antibody images

  • All lanes : Anti-Claudin 3 antibody (ab52231) at 1/500 dilution

    Lane 1 : Extracts from HuvEc cells, untreated.
    Lane 2 : Extracts from HuvEc cells, treated with the immunizing peptide.


    Predicted band size : 23 kDa
    Observed band size : 23 kDa
  • IHC image of ab52231 staining in normal human colon formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab52231, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • ICC/IF image of ab52231 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab52231, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h.Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • All lanes : Anti-Claudin 3 antibody (ab52231)

    Lane 1 : Human urothelial cells (differentiated)
    Lane 2 : Human urothelial cells (non-differentiated)

    Lysates/proteins at 15 µg per lane.

    Secondary
    IRDye® 800-conjugated Goat anti-rabbit IgG polyclonal
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 23 kDa


    Exposure time : 2 minutes

    This image is courtesy of an anonymous Abreview

    See Abreview

  • Claudin 3 was immunoprecipitated using 0.5mg MCF7 Whole cell lysate, 5µg of Rabbit polyclonal to Claudin 3 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, MCF7 Whole cell lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab52231.
    Secondary: Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.
    Band: 18kDa; Claudin 3

References for Anti-Claudin 3 antibody (ab52231)

This product has been referenced in:
  • Han CL  et al. An Informatics-assisted Label-free Approach for Personalized Tissue Membrane Proteomics: Case Study on Colorectal Cancer. Mol Cell Proteomics 10:M110.003087 (2011). WB ; Human . Read more (PubMed: 21209152) »
  • Ye P  et al. CD24 regulated gene expression and distribution of tight junction proteins is associated with altered barrier function in oral epithelial monolayers. BMC Cell Biol 10:2 (2009). Read more (PubMed: 19138432) »

See all 2 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions Reduced Denaturing (4-12% Bis Tris MOPS)
Loading amount 14 µg
Specification Intestinal tissue
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Nov 18 2015

Application Western blot
Sample Rat Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions Reduced Denaturing (4-12% Bis Tris MOPS)
Loading amount 14 µg
Specification Intestinal tissue
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Julien Chevalier

Verified customer

Submitted Oct 01 2015

Application Western blot
Sample Mouse Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions Reduced Denaturing (4-12% Bis Tris MOPS)
Loading amount 14 µg
Specification Intestinal tissue
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Julien Chevalier

Verified customer

Submitted Sep 23 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (human urothelial cells)
Loading amount 15 µg
Specification human urothelial cells
Treatment 5% ABS for 7 days
Gel Running Conditions Reduced Denaturing (4-12% bis tris)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.1% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted May 16 2012

Thank you for submitting the Abreview. This information is very useful both to ourselves and our customers.

In order topublish the Abreviewand activate thediscountcode there are a few details I would like to clarify.Could you please explainbr...

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Thank you for contacting me. I am sorry that the email I sent last Wednesday did not reach you but I'm glad it got through when resent by Kate.

I am glad you would like to test ab52231 for its specificity. I have generated a discount code for...

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Thank you for contacting us and your interest in our products.

I now have more information to share with you in regards to the specificity of the anti-claudin 3 antibody (ab52231).

Through analysis of the immunogen used to raise thi...

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Application Western blot
Sample Cow Cell lysate - whole cell (endothelial cells from cerebral capillaries)
Loading amount 20 µg
Specification endothelial cells from cerebral capillaries
Gel Running Conditions Reduced Denaturing (10 %)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Username

Dr. GOSSELET Fabien

Verified customer

Submitted Apr 17 2008

Thank you for your patience in this matter. The originator of ab52231 (Claudin 3 antibody) has informed us that the immunogenic peptide sequence is not available. I am sorry that I can not provide more information to help with your enquiry.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"