Invitrogen

Villin Polyclonal Antibody

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This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated.

5 Published Figures

View all (27) Villin antibodies

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Villin Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of Villin in methanol-fixed Hep3B cells using a Villin polyclonal antibody (Product # PA5-29078) at a 1:500 dilution.

Villin Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemical analysis of Villin in paraffin-embedded NCI-N87 xenograft using a Villin polyclonal antibody (Product # PA5-29078) at a dilution of 1:500.

Villin Antibody in Western Blot (WB) — Western blot was performed using Anti-Villin Polyclonal Antibody (Product # PA5-29078) and a 92kDa band corresponding to Villin was observed across cell lines and tissues tested. Whole cell extracts (30 µg lysate) of Caco-2 (Lane 1), COLO 205 (Lane 2), HT-29 (Lane 3), Jurkat (Lane 4) and PANC-1 (Lane 5) as seen in Fig (a).Tissue extracts of Mouse Liver (Lane 1), Mouse Colon (Lane 2) and Mouse Jejunum (Lane 3) as seen in Fig (b) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:5000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Villin Antibody in Western Blot (WB) — Western Blot analysis of Villin was performed by separating 50 µg of Mouse tissue extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a Villin Polyclonal Antibody (Product # PA5-29078) at a dilution of 1:1000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Villin Antibody in Western Blot (WB) — Western blot analysis of Villin using 30 µg of H1299 lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a Villin polyclonal antibody (Product # PA5-29078) at a dilution of 1:10,000.

Villin Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target across cell lines and tissues tested owing to their inherent genetic constitution. Relative expression of Villin was observed in High in Caco-2, COLO 205 and HT-29 as compared to low or negative in Jurkat and PANC-1 as seen in Fig (a),Similarly expression was observed high in Mouse Jejunum as compared to low or negative in Mouse Colon and Mouse Liver as seen in Fig (b) using Anti-Villin Polyclonal Antibody (Product # PA5-29078) in Western Blot. (Ref-http://dx.doi.org/10.1016/j.cbpa.2016.08.016, https://doi.org/10.1371/journal.pone.0084221). {RE}

Villin Antibody in Immunocytochemistry (ICC/IF) — FIGURE 7 Intestinal damage and inflammatory responses caused by E. coli on the chip. (A) Conceptual diagram of the microsystem of human inflammatory bowel disease representing co-culture of vascular endothelial cells, enterocyte, macrophages with enterobacteria. (B) The confocal fluorescence views showing the distribution of the tight junction protein - occludin (green) in the epithelial monolayers cultured in the peristaltic human gut-vessel microsystem under control conditions (EC) versus presence of E. coli with (EC + E. coli ) or without ( E. coli ) endothelial cells for 24 h (bar, 50 mum). (C) Quantification of Papp measured by quantitating fluorescent dextran transport across intestinal epithelial layer under the conditions as described in (B) ( n = 3; * P < 0.05, ** p < 0.01, *** P < 0.001). (D) Confocal immunofluorescence views of the sugar chain portion (FITC-WGA, green) of the glycocalyx layer secreted by the Caco2 cells cultured under the conditions described in (B) (nuclei in blue, E. coli in red) (bar, 50 mum). (E) Confocal immunofluorescence views of the villin of the microvilli secreted by the Caco2 cells cultured under the conditions described in (B) (nuclei in blue, villin in red) (bar, 50 mum). Computerized quantification of the coverage (F) and height (G) of the sugar chain portion and villin measured under the conditions described in (B) ( n = 3; ** p < 0.01, *** P < 0.001, **** P < 0.0001). Polarized secretion of proinflammatory cytokines (IL-8, IL-6, TNF

Villin Antibody in Immunocytochemistry (ICC/IF) — FIGURE 8 Anti-inflammatory evaluation of L. casei and antibiotics on the chip. (A) The confocal fluorescence views showing the distribution of the tight junction protein - occludin (green) in the epithelial monolayers cultured in the peristaltic human gut-vessel microsystem with E. coli under control conditions (EC + E. coli ) versus presence of L. casei (EC + E. coli + L. casei ) or P/S (EC + E. coli + P/S) for 24 h (bar, 50 mum). (B) Quantification of Papp measured by quantitating fluorescent dextran transport across intestinal epithelial layer under the conditions described in (A) ( n = 3; * P < 0.05, ** p < 0.01). (C) Confocal immunofluorescence views of the sugar chain portion (FITC-WGA, green) of the glycocalyx layer secreted by the Caco2 cells cultured under the conditions described in (A) (nuclei in blue, E. coli in red) (bar, 50 mum). (D) Confocal immunofluorescence views of the villin of the microvilli layer secreted by the Caco2 cells cultured under the conditions described in (A) (nuclei in blue, villin in red) (bar, 50 mum). Computerized quantification of the coverage (E) and height (F) of the sugar chain portion and villin measured under the conditions described in (A) ( n = 3; ** p < 0.01, *** P < 0.001). Polarized secretion of proinflammatory cytokines (IL-8, IL-6, TNF-alpha, and IL-1beta) in the enteric cavity (H) and vessel lumen (G) under the conditions described in (A) ( n = 3; * P < 0.05, ** p < 0.01).

Villin Antibody in Immunocytochemistry (ICC/IF) — FIGURE 5 Evaluation of intestinal epithelial glycocalyx and microvilli. (A) Confocal immunefluorescence views of the sugar chain portion (FITC-WGA, green) of the glycocalyx layer secreted by the Caco2 cells cultured in the static Transwell system, or in the peristaltic microfluidic chip in the absence (Peris) or presence (Peris + EC) of endothelial cell for 5 days (bar, 50 mum). Computerized quantification of the coverage (B) and height (C) of the sugar chain portion marked by FITC-labeled WGA measured under the conditions described in (A) ( n = 3; * P < 0.05, ** p < 0.01, *** P < 0.001, **** P < 0.0001). (D) Confocal immunofluorescence views of the villin (red) of the microvilli secreted by the Caco2 cells cultured in the static Transwell system, or in the peristaltic microfluidic chip in the absence (Peris) or presence (Peris + EC) of endothelial cell for 5 days (bar, 50 mum). Quantification of the coverage (E) and height (F) of villin done under the conditions as described in (D) ( n = 3; ** p < 0.01, **** P < 0.0001).

Villin Antibody in Immunocytochemistry (ICC/IF) — Published figure using Villin polyclonal antibody (Product # PA5-29078) in Immunocytochemistry

Villin Antibody in Immunocytochemistry (ICC/IF) — Published figure using Villin polyclonal antibody (Product # PA5-29078) in Immunocytochemistry

Product Details

PA5-29078

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500-1:20,000	-
Immunohistochemistry (Paraffin) (IHC (P))	1:100-1:1,000	-
Immunocytochemistry (ICC/IF)	1:100-1:1,000	View 2 publications 2 publications

Product Specifications
Species Reactivity	Human, Mouse, Rat
Published species	Human
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Recombinant fragment corresponding to a region within amino acids 7 and 286 of Human Villin View antigen
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	0.1M tris glycine, pH 7, with 10% glycerol
Contains	0.01% thimerosal
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2546554

Product Specific Information

Recommended positive controls: A431, H1299, HepG2, mouse kidney.

Predicted reactivity: Mouse (89%), Rat (89%), Bovine (90%).

Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Target Information

Villin is part of cytoskeleton associated with the microvillar actin core bundle of gastrointestinal and renal brush border. It can interact with actin in a Ca2+ and phosphoinositide-regulated manner. Villin is a very specific marker for adenocarcinomas of the gastrointestinal tract and the pancreas. It is also expressed in some Merkel cell tumor and adenocarcinoma of the lung, prostate, ovarian and kidney.

⚠WARNING: This product can expose you to chemicals including mercury, which is known to the State of California to cause birth defects or other reproductive harm. For more information go to www.P65Warnings.ca.gov.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Establishment and Application of Peristaltic Human Gut-Vessel Microsystem for Studying Host-Microbial Interaction.

Frontiers in bioengineering and biotechnology

Jing B,Wang ZA,Zhang C,Deng Q,Wei J,Luo Y,Zhang X,Li J,Du Y

PA5-29078 was used in Immunocytochemistry-immunoflourescence to develop a novel gut-vessel microfluidic model to study host-microbial interactions.

Mon Sep 28 00:00:00 EDT 2020

Establishment and Application of Peristaltic Human Gut-Vessel Microsystem for Studying Host-Microbial Interaction.

Frontiers in bioengineering and biotechnology

Jing B,Wang ZA,Zhang C,Deng Q,Wei J,Luo Y,Zhang X,Li J,Du Y

PA5-29078 was used in Immunocytochemistry-immunoflourescence to develop a novel gut-vessel microfluidic model to study host-microbial interactions.

Mon Sep 28 00:00:00 EDT 2020

Modeling radiation injury-induced cell death and countermeasure drug responses in a human Gut-on-a-Chip.

Cell death & disease

Jalili-Firoozinezhad S,Prantil-Baun R,Jiang A,Potla R,Mammoto T,Weaver JC,Ferrante TC,Kim HJ,Cabral JMS,Levy O,Ingber DE

Published figure using Villin polyclonal antibody (Product # PA5-29078) in Immunofluorescence

Wed Feb 14 00:00:00 EST 2018

Bioinformatics

Protein Aliases: OTTHUMP00000207159; Villin-1

Gene Aliases: D2S1471; VIL; VIL1

UniProt ID: (Human) P09327, (Mouse) Q62468

Entrez Gene ID: (Human) 7429, (Mouse) 22349, (Rat) 316521

Molecular Function: non-motor actin binding protein

intestinal D-glucose absorption protein complex assembly apoptotic process epidermal growth factor receptor signaling pathway regulation of cell shape response to bacterium positive regulation of epithelial cell migration actin filament polymerization actin filament depolymerization positive regulation of cell migration positive regulation of actin filament depolymerization epithelial cell differentiation positive regulation of actin filament bundle assembly regulation of microvillus length cellular response to hepatocyte growth factor stimulus positive regulation of multicellular organism growth negative regulation of cysteine-type endopeptidase activity involved in apoptotic process actin nucleation actin filament severing barbed-end actin filament capping regulation of actin nucleation actin filament capping cytoplasmic actin-based contraction involved in cell motility regulation of wound healing cellular response to epidermal growth factor stimulus positive regulation of establishment of protein localization to plasma membrane terminal web assembly regulation of lamellipodium morphogenesis positive regulation of lamellipodium morphogenesis cytoskeleton organization microvillus assembly biological_process

Process(es)

calcium ion binding protein binding phosphatidylinositol-4,5-bisphosphate binding lysophosphatidic acid binding identical protein binding protein homodimerization activity cysteine-type endopeptidase inhibitor activity involved in apoptotic process actin filament binding actin binding metal ion binding molecular_function