概述

  • 产品名称
  • 描述
    兔多克隆抗体to CLCA3
  • 经测试应用
    适用于: IHC-P, WB, ICC/IF, IP, Electron Microscopymore details
  • 种属反应性
    与反应: Mouse
    不与反应: Human
  • 免疫原

    Synthetic peptide:

    EKNHNQEAPNDQNQR

    , corresponding to amino acids 253-267 of Mouse CLCA3

性能

应用

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.

应用 Ab评论 说明
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.

靶标

  • 相关性
    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.
  • 细胞定位
    Secreted
  • 数据库链接
  • 别名
    • 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

图片

  • 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.

文献

This product has been referenced in:
  • 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) »
  • Mushaben EM  et al. Differential effects of rapamycin and dexamethasone in mouse models of established allergic asthma. PLoS One 8:e54426 (2013). WB ; Mouse . Read more (PubMed: 23349887) »

See all 7 Publications for this product

客户评价及客户问答

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
Username

Abcam user community

Verified customer

提交于 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
Username

Abcam user community

Verified customer

提交于 Mar 01 2012

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 a...

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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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