Recombinant Anti-Villin antibody [SP145] (ab130751)
- Datasheet
- References (5)
- Protocols
Overview
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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
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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.
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General notes
This product is a recombinant rabbit monoclonal antibody.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.6
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA -
Concentration information loading... -
Purity
Protein A/G purified -
Purification notes
Purified from TCS by protein A/G. -
Clonality
Monoclonal -
Clone number
SP145 -
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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Positive Controls
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Recombinant Protein
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.
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| 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
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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
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Database links
- Entrez Gene: 396423 Chicken
- Entrez Gene: 7429 Human
- Entrez Gene: 22349 Mouse
- Omim: 193040 Human
- SwissProt: P02640 Chicken
- SwissProt: P09327 Human
- SwissProt: Q62468 Mouse
- Unigene: 654595 Human
see all -
Alternative names
- D2S1471 antibody
- OTTHUMP00000164145 antibody
- VIL antibody
see all
Images
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Villin antibody [SP145] (ab130751)](https://www.abcam.com/ps/products/130/ab130751/Images/ab130751-145088-anti-villin-antibody-sp145-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Villin antibody [SP145] (ab130751)ab130751, at 1/100 dilution, staining Villin in formalin-fixed, paraffin-embedded Human colon tissue by Immunohistochemistry. -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Villin antibody [SP145] (ab130751)](https://www.abcam.com/ps/products/130/ab130751/Images/ab130751-227994-anti-villin-antibody-sp145-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Villin antibody [SP145] (ab130751)This image is courtesy of an anonymous Abreviewab130751 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.
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![Immunoprecipitation - Anti-Villin antibody [SP145] (ab130751)](https://www.abcam.com/ps/products/130/ab130751/Images/ab130751-234720-anti-villin-antibody-sp145-immunoprecipitation.jpg)
Immunoprecipitation - Anti-Villin antibody [SP145] (ab130751)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
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![Flow Cytometry - Anti-Villin antibody [SP145] (ab130751)](https://www.abcam.com/ps/products/130/ab130751/Images/ab130751-204531-anti-villin-antibody-sp145-flow-cytometry.jpg)
Flow Cytometry - Anti-Villin antibody [SP145] (ab130751)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. -
![Western blot - Anti-Villin antibody [SP145] (ab130751)](https://www.abcam.com/ps/products/130/ab130751/Images/ab130751-145087-anti-villin-antibody-sp145-western-blot.jpg)
Western blot - Anti-Villin antibody [SP145] (ab130751)Anti-Villin antibody [SP145] (ab130751) at 1/100 dilution + HT-29 cell lysate
Predicted band size: 93 kDa
Protocols
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) »
