Overview

  • Product name
  • Description
    Rabbit polyclonal to CLCA3
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, WB, ICC/IF, IP, Electron Microscopymore details
  • Species reactivity
    Reacts with: Mouse
    Does not react with: Human
  • Immunogen

    Synthetic peptide:

    EKNHNQEAPNDQNQR

    , corresponding to amino acids 253-267 of Mouse CLCA3

Properties

Applications

Our Abpromise guarantee covers the use of ab46512 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
IHC-P 1/8000. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
WB 1/1000. Predicted molecular weight: 100 kDa.
ICC/IF 1/1000.
IP 1/100 - 1/500.
Electron Microscopy 1/2000.

Target

  • Relevance
    mCLCA3 (alias gob-5) is the third murine member of the family of calcium-activated chloride channels (CLCA-family). mCLCA3 plays a role in diseases with secretory dysfunctions, including asthma and cystic fibrosis. The mCLCA3 protein is located in mucin granule membranes of goblet cells of the intestinal, respiratory and reproductive tracts. It is also secreted into the mucin layer on mucosal membranes. The protein appears to be involved in the synthesis, condensation or secretion of mucins.
  • Cellular localization
    Secreted
  • Database links
  • Alternative names
    • Calcium-activated chloride channel regulator family member 3 antibody
    • Chloride channel accessory 3 pseudogene antibody
    • chloride channel calcium activated 3 antibody
    • Clca1 antibody
    • Clca2 antibody
    • CLCA3P antibody
    • Gob 5 antibody
    • gob5 antibody
    • hCLCA3 antibody
    • mCLCA3 antibody
    see all

Images

  • Formalin-fixed, paraffin-embedded mouse colon tissue sections stained with anti-CLCA3 ab46512 used at 1/8000 dilution.
  • Anti-CLCA3 antibody (ab46512) at 1/500 dilution + MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 100 kDa
    Observed band size: 35 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 16 kDa, 95 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 1 minute


    CLCA3 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
  • ICC/IF image of ab46512 stained MEF1 cells. The cells were 4% PFA fixed (10mins) 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 (ab46512, 1/200 dilution) 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.

References

This product has been referenced in:
  • Acciani TH  et al. Epidermal growth factor receptor signalling regulates granulocyte-macrophage colony-stimulating factor production by airway epithelial cells and established allergic airway disease. Clin Exp Allergy 46:317-28 (2016). Read more (PubMed: 26263242) »
  • Erickson NA  et al. The Goblet Cell Protein Clca1 (Alias mClca3 or Gob-5) Is Not Required for Intestinal Mucus Synthesis, Structure and Barrier Function in Naive or DSS-Challenged Mice. PLoS One 10:e0131991 (2015). IF . Read more (PubMed: 26162072) »
See all 8 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

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 (Lung)
Specification
Lung
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH6.0
Permeabilization
Yes - Methanol and 30% Hydrogen peroxide
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Mar 19 2012

Application
Western blot
Sample
Rabbit Tissue lysate - whole (Lung)
Loading amount
60 µg
Specification
Lung
Gel Running Conditions
Reduced Non-Denaturing (Native) (4-12%)
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 Mar 01 2012

Answer

Thank you very much for your interest in ab46512.  To our knowledge, ab46512 has not been tested in frozen tissue sections (IHC-Fr).  There is no reason that I can see that it wouldn't work, but since we haven't tested it in IHC-Fr we wouldn't be able to guarantee it. Therefore, I can offer a discount off a future purchase if you buy ab46512 now, test it in IHC-Fr and 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 ab46512 in IHC-Fr. I will then send a discount code. This code must be issued before purchasing ab46512 so please wait for my reply before ordering. 2. Purchase ab46512 either by phone, fax, or online (www.abcam.com). 3. Test it in IHC-Fr. 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 ab46512 turns out to be unsuitable for IHC-Fr, 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: www.abcam.com/collaborationdiscount.  

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

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