Overview

  • Product name

    Anti-Claudin 1 antibody [EPRR18871]
    See all Claudin 1 primary antibodies
  • Description

    Rabbit monoclonal [EPRR18871] to Claudin 1
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Claudin 1 aa 150 to the C-terminus. The exact sequence is proprietary.
    Database link: O95832

  • Positive control

    • WB: HEK-293, A431 and HepG2 whole cell lysates; human fetal liver and fetal kidney lysates; human Claudin 1 full length recombinant protein. IHC-P: Human skin, cervix, colon cancer and lung squamous cell cancer tissues. ICC/IF: A431 and HepG2 cells. Flow Cyt: HepG2 cells. IP: HepG2 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

  • 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

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPRR18871
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab211737 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
ICC/IF 1/1000.
Flow Cyt 1/60.
IP 1/40.
IHC-P 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/2000. Detects a band of approximately 18 kDa (predicted molecular weight: 22 kDa).

Target

  • Function

    Claudins function as major constituents of the tight junction complexes that regulate the permeability of epithelia. While some claudin family members play essential roles in the formation of impermeable barriers, others mediate the permeability to ions and small molecules. Often, several claudin family members are coexpressed and interact with each other, and this determines the overall permeability. CLDN1 is required to prevent the paracellular diffusion of small molecules through tight junctions in the epidermis and is required for the normal barrier function of the skin. Required for normal water homeostasis and to prevent excessive water loss through the skin, probably via an indirect effect on the expression levels of other proteins, since CLDN1 itself seems to be dispensable for water barrier formation in keratinocyte tight junctions (PubMed:23407391).
    (Microbial infection) Acts as a receptor for hepatitis C virus in hepatocytes (PubMed:17325668). Acts as a receptor for dengue virus (PubMed:24074594).
  • Tissue specificity

    Strongly expressed in liver and kidney. Expressed in heart, brain, spleen, lung and testis.
  • Involvement in disease

    Ichthyosis-sclerosing cholangitis neonatal syndrome
  • Sequence similarities

    Belongs to the claudin family.
  • Cellular localization

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

  • Alternative names

    • Claudin-1 antibody
    • Claudin1 antibody
    • CLD 1 antibody
    • CLD1 antibody
    • CLD1_HUMAN antibody
    • CLDN 1 antibody
    • Cldn1 antibody
    • ILVASC antibody
    • SEMP 1 antibody
    • SEMP1 antibody
    • Senescence associated epithelial membrane protein 1 antibody
    • Senescence associated epithelial membrane protein antibody
    • Senescence-associated epithelial membrane protein antibody
    see all

Images

  • All lanes : Anti-Claudin 1 antibody [EPRR18871] (ab211737) at 1/2000 dilution

    Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 2 : A431 (Human epidermoid carcinoma cell line) whole cell lysate
    Lane 3 : HepG2 (Human liver hepatocellular carcinoma 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: 22 kDa
    Observed band size: 18 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1/2: 15 seconds; Lane 3: 5 seconds.

    The expression profile/ molecular weight  observed is consistent with what has been described in the literature PMID: 26601075.

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling Claudin 1 with ab211737 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membrane staining on human colon 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.

  • All lanes : Anti-Claudin 1 antibody [EPRR18871] (ab211737) at 1/2000 dilution

    Lane 1 : Human fetal liver lysate
    Lane 2 : 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 1/10000 dilution

    Predicted band size: 22 kDa
    Observed band size: 18 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile/ molecular weight  observed is consistent with what has been described in the literature PMID: 26601075.

  • Anti-Claudin 1 antibody [EPRR18871] (ab211737) at 1/5000 dilution + Human Claudin 1 full length recombinant protein at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 22 kDa
    Observed band size: 45 kDa why is the actual band size different from the predicted?


    Exposure time: 1 second


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Human Claudin 1 full length recombinant protein contains aa1-211 with His-Tag® and GST-tag. This recombinant was made in house.

  • Immunohistochemical analysis of paraffin-embedded human skin tissue labeling Claudin 1 with ab211737 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membrane staining on human skin squamous cells 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 cervix tissue labeling Claudin 1 with ab211737 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membrane staining on human cervix 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 lung squamous cell cancer tissue labeling Claudin 1 with ab211737 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membrane staining on human lung squamous cell 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.

  • Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma tissue labeling Claudin 1 with ab211737 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Negative control: Negative staining on human lung adenocarcinoma, which is consistent with the literature (PMID: 17585317). 

    Counter stained with Hematoxylin.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A431 (Human epidermoid carcinoma cell line) cells labeling Claudin 1 with ab211737 at 1/1000 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 A431 cell line.The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab211737 at 1/1000 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.

    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Claudin 1 with ab211737 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing membranous staining on HepG2 cell line. 

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab211737 at 1/1000 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.

    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Claudin 1 with ab211737 at 1/60 dilution (red) compared with a rabbit monoclonal IgG 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 1 was immunoprecipitated from 0.35 mg of HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate with ab211737 at 1/40 dilution.

    Western blot was performed from the immunoprecipitate using ab211737 at 1/2000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

    Lane 1: HepG2 whole cell lysate 10µg (Input).

    Lane 2: ab211737 IP in HepG2 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab211737 in HepG2 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 seconds.

References

This product has been referenced in:

  • Feng L  et al. Bisphenol A increases intestinal permeability through disrupting intestinal barrier function in mice. Environ Pollut 254:112960 (2019). Read more (PubMed: 31394344) »
  • Hu J  et al. Lactobacillus frumenti Facilitates Intestinal Epithelial Barrier Function Maintenance in Early-Weaned Piglets. Front Microbiol 9:897 (2018). Read more (PubMed: 29867808) »
See all 2 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Skin)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris pH9
Permeabilization
No
Specification
Skin
Blocking step
Casein as blocking agent for 10 minute(s) · Concentration: 1%
Fixative
Formaldehyde

Dr. Faye Doherty

Verified customer

Submitted Jun 05 2019

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up