Recombinant Anti-Villin antibody [SP145] (ab130751)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [SP145] to Villin
- Suitable for: IP, mIHC, WB, IHC-P, Flow Cyt (Intra)
- Reacts with: Human
Related conjugates and formulations
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: IP, mIHC, WB, IHC-P, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, 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. mIHC: Human colon tissue. Flow cyto(intra): HT-29 cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.60
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA -
Concentration information loading...
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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
Applications
The Abpromise guarantee
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 |
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IP |
Use a concentration of 5 µg/ml.
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mIHC |
Use at an assay dependent concentration.
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WB |
1/100. Predicted molecular weight: 93 kDa.
Incubate for 1 hour at room temperature. |
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IHC-P | (3) |
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. |
Flow Cyt (Intra) |
1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Notes |
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IP
Use a concentration of 5 µg/ml. |
mIHC
Use at an assay dependent concentration. |
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. |
Flow Cyt (Intra)
1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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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Flow cytometry overlay histogram showing left HT-29 positive cells and right negative PANC-1 stained with ab130751 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab130751) (1x 106 in 100μl at 0.2μg/ml (1/11200)) for 30min at 22°C.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
This antibody gave a positive signal in HT-29 Fixed with 4% formaldehyde (10 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.
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Fluorescence multiplex immunohistochemical analysis of the human colon (Formalin/PFA-fixed paraffin-embedded sections). Panel A: merged staining of anti-Villin (ab245749, gray; Opal™690), anti-liver FABP (ab240401, green; Opal™520) and anti-MUC2 (ab272692, red; Opal™570) on human colon. Panel B: anti-liver FABP stained on enterocytes. Panel C: anti-Villin stained on apical border. Panel D: anti-MUC2 stained on goblet cells. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining: in the order of ab245749 (1/1000 dilution), ab240401 (1/8000 dilution), and ab272692 (1/5000 dilution) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.
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ab130751, at 1/100 dilution, staining Villin in formalin-fixed, paraffin-embedded Human colon tissue by Immunohistochemistry.
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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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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.
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Anti-Villin antibody [SP145] (ab130751) at 1/100 dilution + HT-29 cell lysate
Predicted band size: 93 kDa
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (39)
ab130751 has been referenced in 39 publications.
- Sanson R et al. Axin1 Protects Colon Carcinogenesis by an Immune-Mediated Effect. Cell Mol Gastroenterol Hepatol 15:689-715 (2023). PubMed: 36356835
- Ren J et al. Sex Differences of Radiation Damage in High-Fat-Diet-Fed Mice and the Regulatory Effect of Melatonin. Nutrients 15:N/A (2022). PubMed: 36615722
- Centonze M et al. The Fast Track for Intestinal Tumor Cell Differentiation and In Vitro Intestinal Models by Inorganic Topographic Surfaces. Pharmaceutics 14:N/A (2022). PubMed: 35057113
- Saari J et al. Toward Xeno-Free Differentiation of Human Induced Pluripotent Stem Cell-Derived Small Intestinal Epithelial Cells. Int J Mol Sci 23:N/A (2022). PubMed: 35163236
- Reicher N et al. Intra-amniotic administration of l-glutamine promotes intestinal maturation and enteroendocrine stimulation in chick embryos. Sci Rep 12:2645 (2022). PubMed: 35173228