Overview

  • Product name

    Anti-Villin antibody [SP145]
    See all Villin primary antibodies
  • Description

    Rabbit monoclonal [SP145] to Villin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, IP, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Chicken, Cow, Pig
  • Immunogen

    Synthetic peptide within Human Villin aa 800 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P09327

  • Positive control

    • Human colon, small intestine and kidney tissues; HT-29 cell lysate.
  • General notes

     

     

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab130751 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
Flow Cyt 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

IP Use a concentration of 5 µg/ml.
WB 1/100. Predicted molecular weight: 93 kDa. Incubate for 1 hour at room temperature.
IHC-P 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Primary incubation for 10 minutes at room temperature.

Target

  • Function

    Ca(2+)-regulated actin-binding protein.
  • Tissue specificity

    Major component of microvilli of intestinal epithelial cells and kidney proximal tubule cells.
  • Sequence similarities

    Belongs to the villin/gelsolin family.
    Contains 6 gelsolin-like repeats.
    Contains 1 HP (headpiece) domain.
  • Domain

    Consists of a large core fragment, the N-terminal portion, and a small headpiece, the C-terminal portion. The headpiece binds F-actin strongly in both the presence and absence of calcium.
  • Cellular localization

    Cytoplasm > cytoskeleton.
  • Information by UniProt
  • Database links

  • Alternative names

    • D2S1471 antibody
    • OTTHUMP00000164145 antibody
    • VIL antibody
    • VIL1 antibody
    • VILI_HUMAN antibody
    • Villin 1 antibody
    • Villin-1 antibody
    • Villin1 antibody
    see all

Images

  • ab130751, at 1/100 dilution, staining Villin in formalin-fixed, paraffin-embedded Human colon tissue by Immunohistochemistry.
  • ab130751 staining Villin in mouse intestine tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 5% serum for 30 minutes at 25°C; antigen retrieval was by heat mediation in 10mM citrate buffer, pH 6. Samples were incubated with primary antibody (1/1000) for 16 hours at 4°C. An Alexa Fluor® 568-conjugated goat anti-rabbit IgG monoclonal (1/250) was used as the secondary antibody.

    See Abreview

  • Villin was immunoprecipitated using 0.5mg SW480 whole cell extract, 5µg of Rabbit polyclonal to Villin and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, SW480 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab130751.

    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).

    Band: 93kDa; Villin

  • Overlay histogram showing Caco 2 cells stained with ab130751 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab130751, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
  • Anti-Villin antibody [SP145] (ab130751) at 1/100 dilution + HT-29 cell lysate

    Predicted band size: 93 kDa

References

This product has been referenced in:

  • Jones JC  et al. Cellular Plasticity of Defa4Cre-Expressing Paneth Cells in Response to Notch Activation and Intestinal Injury. Cell Mol Gastroenterol Hepatol 7:533-554 (2019). Read more (PubMed: 30827941) »
  • Koren E  et al. ARTS mediates apoptosis and regeneration of the intestinal stem cell niche. Nat Commun 9:4582 (2018). Read more (PubMed: 30389919) »
See all 5 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Small Intestine)
Antigen retrieval step
Heat mediated
Permeabilization
No
Specification
Small Intestine
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jul 27 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM citrate buffer pH6
Sample
Mouse Tissue (intestine)
Specification
intestine
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jun 13 2014

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