Recombinant Anti-Claudin 4 antibody [EPRR17575] (ab210796)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPRR17575] to Claudin 4
- Suitable for: IHC-P, WB, IP
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-Claudin 4 antibody [EPRR17575]
See all Claudin 4 primary antibodies -
Description
Rabbit monoclonal [EPRR17575] to Claudin 4 -
Host species
Rabbit -
Specificity
The mouse recommendation is based on the WB results. We do not guarantee IHC-P for mouse.
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Tested applications
Suitable for: IHC-P, WB, IPmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: LNCaP, PC-3, SW480, MCF7, SK-OV-3, NIH:OVCAR-3, 4T1. IHC-P: Human colon, endometrium, liver, cholangiocarcinoma and breast cancer tissues. IP: SW480 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
EPRR17575 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab210796 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 |
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IHC-P | (2) |
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/1000. Detects a band of approximately 18 kDa (predicted molecular weight: 22 kDa).
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IP |
1/30.
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Notes |
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IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Detects a band of approximately 18 kDa (predicted molecular weight: 22 kDa). |
IP
1/30. |
Target
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Function
Plays a major role in tight junction-specific obliteration of the intercellular space. -
Involvement in disease
Note=CLDN4 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
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Database links
- Entrez Gene: 1364 Human
- Entrez Gene: 12740 Mouse
- Omim: 602909 Human
- SwissProt: O14493 Human
- SwissProt: O35054 Mouse
- Unigene: 647036 Human
- Unigene: 699253 Human
- Unigene: 7339 Mouse
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Alternative names
- Claudin-4 antibody
- Claudin4 antibody
- CLD4_HUMAN antibody
see all
Images
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All lanes : Anti-Claudin 4 antibody [EPRR17575] (ab210796) at 1/1000 dilution
Lane 1 : Wild-type MCF7 cell lysate
Lane 2 : CLDN4 knockout MCF7 cell lysate
Lane 3 : PC-3 cell lysate
Lane 4 : U-2 OS cell lysate
Lane 5 : SW480 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 22 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Claudin 4 antibody [EPRR17575] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab210796 was shown to bind specifically to Claudin 4. A band was observed at 17 kDa in wild-type MCF7 cell lysates with no signal observed at this size in CLDN4 knockout cell line ab274946 (knockout cell lysate ab275004). To generate this image, wild-type and CLDN4 knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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Anti-Claudin 4 antibody [EPRR17575] (ab210796) at 1/1000 dilution + 4T1 (mouse mammary gland carcinoma epithelial cell), whole cell lysate at 20 µg
Secondary
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?
Exposure time: 3 minutesBlocking and diluting buffer and concentration: 5% NFDM/TBST
The identity of the higher MW band at approximately 35 kDa is unknown.
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Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Claudin 4 with ab210796 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Membrane and staining on epithelial cells of human colon 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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Lanes 1-2 : Anti-Claudin 4 antibody [EPRR17575] (ab210796) at 1/2000 dilution
Lanes 3-6 : Anti-Claudin 4 antibody [EPRR17575] (ab210796) at 1/1000 dilution
Lane 1 : LNCaP (Human prostate cancer cell line) whole cell lysate
Lane 2 : PC-3 (Human prostate adenocarcinoma cell line) whole cell lysate
Lane 3 : SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate
Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lane 5 : SK-OV-3 (Human ovarian cancer cell line) whole cell lysate
Lane 6 : NIH:OVCAR-3 (Human ovary 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: 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 1/2/3/6: 1 minute; Lane 4: 30 seconds; Lane 5: 3 minutes.
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Immunohistochemical analysis of paraffin-embedded human endometrium tissue labeling Claudin 4 with ab210796 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Membrane staining on epithelial cells of human endometrium 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 liver tissue labeling Claudin 4 with ab210796 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Weak membrane staining on biliary epithelium of human liver 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 cholangiocarcinoma tissue labeling Claudin 4 with ab210796 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Membrane staining on human cholangiocarcinoma 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 hepatocellular carcinoma tissue labeling Claudin 4 with ab210796 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Negative staining on human hepatocellular carcinoma.
Counter stained with Hematoxylin.
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 4 with ab210796 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Membrane staining on epithelial 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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Claudin 4 was immunoprecipitated from 1mg of SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate with ab210796 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab210796 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: SW480 whole cell lysate 10µg (Input).
Lane 2: ab210796 IP in SW480 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab210796 in SW480 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (11)
ab210796 has been referenced in 11 publications.
- Ma Y et al. Dietary supplementation of thiamine enhances colonic integrity and modulates mucosal inflammation injury in goats challenged by lipopolysaccharide and low pH. Br J Nutr N/A:1-11 (2022). PubMed: 35057872
- Brice DP et al. Interleukin-27 Regulates the Function of the Gastrointestinal Epithelial Barrier in a Human Tissue-Derived Organoid Model. Biology (Basel) 11:N/A (2022). PubMed: 35336801
- Sun X et al. Improvement of the bladder perfusion curative effect through tight junction protein degradation induced by magnetic temperature-sensitive hydrogels. Front Bioeng Biotechnol 10:958072 (2022). PubMed: 35992356
- De Luccia TPB et al. The effect of Gestational Diabetes Mellitus on the fetal compartment. J Reprod Immunol 145:103314 (2021). PubMed: 33836321
- Ma Y et al. Thiamine Alleviates High-Concentrate-Diet-Induced Oxidative Stress, Apoptosis, and Protects the Rumen Epithelial Barrier Function in Goats. Front Vet Sci 8:663698 (2021). PubMed: 34095275