Overview

  • Product name
  • Description
    Rabbit polyclonal to Claudin 3
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Human, Pig
  • Immunogen

    Synthetic peptide within Mouse Claudin 3 aa 200 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: Q9Z0G9

  • Positive control
    • Tested with small intestine cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab15102 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 1/100.
WB Use at an assay dependent concentration. PubMed: 18192323
IHC-P 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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

  • ICC/IF image of ab15102 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 (ab15102, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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.

  • ab15102 (2µg/ml) staining Claudin 3 in human ileum using an automated system (DAKO Autostainer Plus). Using this protocol there is strong membrane staining of the mucosal epithelial cells.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • Immunocytochemistry/ Immunofluorescence analysis of pig primary porcine brain capillary endothelial cells labeling Claudin 3 with ab15102 (undiluted). Cells were fixed with Acetone/Methanol (3:7) and blocked with 10% BSA for 30 minutes at 22°C. A Goat anti-Rabbit IgG (H+L) Alexa Fluor® 488 conjugate secondary antibody was used at 1/200. DAPI nuclear staining.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling Claudin 3 with ab15102 at 1/300 dilution. Slides were rehydrated and then boiled for antigen retrieval in TE pH 9 buffer for 15min in standard microwave oven at 650W. After 30min blocking in PBS-Triton 0,1% / BSA 5% slides were incubated overnight with Anti-Claudin 3 antibody (ab15102) at 1/300 dilution at 4°C. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/200 dilution for 1h. Nuclei were stained with Hoechst.

    See Abreview

  • Immunohistochemical analysis of PFA-fixed, paraffin embedded BALB/c murine ear tissue sections.

    Claudin 3 was stained, using ab15102 at 1/10 dilution, in the inner ear at the organ of Corti, Reissner’s membrane, periosteal lining of otic capsule, and the apical surface of the marginal cells of stria vascularis (upper panel) and in the middle ear intercellular junctions along lateral and basal cell membranes in the mucosal epithelium (lower panel). An AlexaFluor®488-conjugated IgG (1/50) was used as the secondary antibody.

References

This product has been referenced in:
  • Tokuhara Y  et al. Nuclear expression of claudin-3 in human colorectal adenocarcinoma cell lines and tissues. Oncol Lett 15:99-108 (2018). Read more (PubMed: 29285188) »
  • Zeniya S  et al. Angubindin-1 opens the blood-brain barrier in vivo for delivery of antisense oligonucleotide to the central nervous system. J Control Release 283:126-134 (2018). ICC/IF ; Rat . Read more (PubMed: 29753959) »
See all 19 Publications for this product

Customer reviews and Q&As

1-10 of 13 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (Mature Porcine Jejunum)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA pH 6.0
Permeabilization
No
Specification
Mature Porcine Jejunum
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Formaldehyde

Alex Pasternak

Verified customer

Submitted Aug 26 2016

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

Abcam user community

Verified customer

Submitted Sep 07 2015

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

Abcam user community

Verified customer

Submitted Aug 04 2015

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

Abcam user community

Verified customer

Submitted Jul 29 2015

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Non-reduced Denaturing (15%)
Sample
Mouse Tissue lysate - whole (lung)
Specification
lung
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Apr 01 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Sample
Chicken Cell (neural fold stage)
Specification
neural fold stage
Permeabilization
Yes - 0.3% tritonX-100 in PBS
Fixative
10% trichloroacetic acid

Ms. Amanda Baumholtz

Verified customer

Submitted Jun 25 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Liver)
Specification
Liver
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH 9.0
Permeabilization
Yes - PBS-Triton 0,1% 10min
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Daniel Alpern

Verified customer

Submitted Aug 06 2012

Answer

Thank you for contacting us. We have tested this antibody only in ICC/IF, WB and IHC-Phowevernot inflow cytometry. This antibody can be used in Flow Cytometry however due to lack of tested data we are unable to confirm it. Therefore, I can offer a discount off a future purchase if you buy ab15102 now, test it inFlow Cytometryand submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free (PRIMARY ANTIBODY) If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and test ab15102 in Flow Cyto. I will then send a discount code. This code must be issued before purchasing ab15102 so please wait for my reply before ordering. 2. Purchase ab15102 either by phone, fax, or online (www.abcam.com). 3. Test it in Flow Cyto. 4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews. 5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any [PRIMARY ANTIBODY) ordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab15102 turns out to be unsuitable for Flow Cyto, you will still receive the discount on your next purchase after your Abreview has been submitted. Please let me know if you have any questions about this offer and I would be happy to help you further. The Terms and Conditions of this offer can be found at: https://www.abcam.com/collaborationdiscount. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. PS: We recommend permeabilizing the cell when using this antibody in Flow cytometry.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Pig Cell (primary porcine brain capillary endothelial cells)
Specification
primary porcine brain capillary endothelial cells
Fixative
Acetone/Methanol (3:7)
Permeabilization
No
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C

Mrs. Julia Stab

Verified customer

Submitted Oct 14 2011

Answer

Thank you for your enquiry and I'm sorry to hear that you are experiencing difficulty with ab15102. This antibody hasn't previously been tested for application in Western blotting, so we don't know if it detects claudin 3 under these conditions. The antibody was characterized for application in IHC-formalin-fixed paraffin-embedded sections with small intestine cells. I would suggest that you run a positive control with the antibody; I found in SwissProt that claudin 3 is expressed at high levels in liver and lung and, at lower levels, in kidney and testis so you may want to try a liver or lung lysate. The predicted MW of claudin 3 is approximately 23 kDa and you're seeing a band at approximately 30 kDa, which is pretty close. The protein sample may have multiple modified forms (such as glycosylation, etc) so check the literature to see and check if there are any reports of variants. Also, I think that you can reduce the amount of lysate that you are loading t0 30 ug, which should be sufficient. I do suggest that you submit an Abreview. We always encourage customers to feed their results back to us, whether positive or negative, and make all information about each of our products available to researchers. If you send us an Abreview, we can offer you 50 Abcam Points (a further 100 Abpoints will be offered for an image). To find out more about our Loyalty Scheme and Abpoints, please visit our website and look at Your Account panel (left bottom section on our home page) or the Abreviews section on the product datasheet. You can submit the Abreview if you click on the Abreview section (top menu bar) on the on-line datasheet of this antibody. It is a very straightforward process. Should you need any help how to submit an Abreview, then please do not hesitate to contact us again. I hope this helps. Please contact us again if you have any additional questions.

Read More

1-10 of 13 Abreviews or Q&A

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