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Villin1 Recombinant Rabbit Monoclonal Antibody [JE39-08]

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Specification

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Catalog# ET7110-55

Villin1 Recombinant Rabbit Monoclonal Antibody [JE39-08]

Application

  • WB

  • IHC-P

  • IP

  • IF-Tissue

Reactivity

  • Human

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Villin1 Recombinant Rabbit Monoclonal Antibody [JE39-08]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human Villin1 aa 661-710 / 827.

Species Reactivity

Human

Validated Applications

WB, IHC-P, IP, IF-Tissue

Molecular Weight

Predicted band size: 93 kDa

Positive Control

Caco-2 cell lysate, COLO205 cell lysate, HepG2 cell lysate, AGS cell lysate, human colon tissue, human colon cancer tissue, human kidney tissue.

Conjugation

unconjugated

Clone Number

JE39-08

RRID

AB_3071019

Reactivity Data

WB IHC-P IP IF-Tissue
Human
Tested Tested
Tested Tested
Tested Tested
Tested Tested
Mouse

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IHC-P

  • 1:1,000

  • IP

  • 1-2μg/sample

  • IF-Tissue

  • 1:200

Target

Function

This gene encodes a member of a family of calcium-regulated actin-binding proteins. This protein represents a dominant part of the brush border cytoskeleton which functions in the capping, severing, and bundling of actin filaments. Two mRNAs of 2.7 kb and 3.5 kb have been observed; they result from utilization of alternate poly-adenylation signals present in the terminal exon. Epithelial cell-specific Ca2+-regulated actin-modifying protein that modulates the reorganization of microvillar actin filaments. Plays a role in the actin nucleation, actin filament bundle assembly, actin filament capping and severing. Binds phosphatidylinositol 4,5-bisphosphate (PIP2) and lysophosphatidic acid (LPA); binds LPA with higher affinity than PIP2. Binding to LPA increases its phosphorylation by SRC and inhibits all actin-modifying activities. Binding to PIP2 inhibits actin-capping and -severing activities but enhances actin-bundling activity. Regulates the intestinal epithelial cell morphology, cell invasion, cell migration and apoptosis. Protects against apoptosis induced by dextran sodium sulfate (DSS) in the gastrointestinal epithelium. Appears to regulate cell death by maintaining mitochondrial integrity. Enhances hepatocyte growth factor (HGF)-induced epithelial cell motility, chemotaxis and wound repair. Upon S.flexneri cell infection, its actin-severing activity enhances actin-based motility of the bacteria and plays a role during the dissemination.

Background References

1. Miears HL. et. al. Plant Villin Headpiece Domain Demonstrates a Novel Surface Charge Pattern and High Affinity for F-Actin. Biochemistry. 2018 Mar 20;57(11):1690-1701.

2. Li Z. et. al. Dual Stain With SATB2 and CK20/Villin Is Useful to Distinguish Colorectal Carcinomas From Other Tumors. Am J Clin Pathol. 2018 Feb 17;149(3):241-246.

Sequence Similarity

Belongs to the villin/gelsolin family.

Tissue Specificity

Specifically expressed in epithelial cells. Major component of microvilli of intestinal epithelial cells and kidney proximal tubule cells. Expressed in canalicular microvilli of hepatocytes (at protein level).

Post-translational Modification

Tyrosine phosphorylation is induced by epidermal growth factor (EGF) and stimulates cell migration (By similarity). Phosphorylated on tyrosine residues by SRC. The unphosphorylated form increases the initial rate of actin-nucleating activity, whereas the tyrosine-phosphorylated form inhibits actin-nucleating activity, enhances actin-bundling activity and enhances actin-severing activity by reducing high Ca(2+) requirements. The tyrosine-phosphorylated form does not regulate actin-capping activity. Tyrosine phosphorylation is essential for cell migration: tyrosine phosphorylation sites in the N-terminus half regulate actin reorganization and cell morphology, whereas tyrosine phosphorylation sites in the C-terminus half regulate cell migration via interaction with PLCG1.

Subcellular Location

Cell projection, Cytoplasm, Cytoskeleton.

UNIPROT

SwissProt: P09327 Human

Synonyms

D2S1471 antibody

OTTHUMP00000164145 antibody

VIL antibody

VIL1 antibody

VILI_HUMAN antibody

Villin 1 antibody

Villin-1 antibody

Villin1 antibody

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of Villin1 on different lysates with Rabbit anti-Villin1 antibody (<a href="/products/ET7110-55" style="font-weight: bold;text-decoration: underline;">ET7110-55</a>) at 1/5,000 dilution.<br /><br />Lane 1: Caco-2 cell lysate (20 µg/Lane)<br />Lane 2: COLO205 cell lysate (20 µg/Lane)<br />Lane 3: HepG2 cell lysate (20 µg/Lane)<br />Lane 4: AGS cell lysate (20 µg/Lane)<br />Lane 5: PANC-1 cell lysate (negative) (20 µg/Lane)<br /><br />Predicted band size: 93 kDa<br />Observed band size: 93 kDa<br /><br />Exposure time: 10 seconds; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET7110-55" style="font-weight: bold;text-decoration: underline;">ET7110-55</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of Villin1 on different lysates with Rabbit anti-Villin1 antibody (ET7110-55) at 1/5,000 dilution.

    Lane 1: Caco-2 cell lysate (20 µg/Lane)
    Lane 2: COLO205 cell lysate (20 µg/Lane)
    Lane 3: HepG2 cell lysate (20 µg/Lane)
    Lane 4: AGS cell lysate (20 µg/Lane)
    Lane 5: PANC-1 cell lysate (negative) (20 µg/Lane)

    Predicted band size: 93 kDa
    Observed band size: 93 kDa

    Exposure time: 10 seconds; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7110-55) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-Villin1 antibody (<a href="/products/ET7110-55" style="font-weight: bold;text-decoration: underline;">ET7110-55</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7110-55" style="font-weight: bold;text-decoration: underline;">ET7110-55</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-Villin1 antibody (ET7110-55) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-55) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-Villin1 antibody (<a href="/products/ET7110-55" style="font-weight: bold;text-decoration: underline;">ET7110-55</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7110-55" style="font-weight: bold;text-decoration: underline;">ET7110-55</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-Villin1 antibody (ET7110-55) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-55) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Villin1 antibody (<a href="/products/ET7110-55" style="font-weight: bold;text-decoration: underline;">ET7110-55</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7110-55" style="font-weight: bold;text-decoration: underline;">ET7110-55</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Villin1 antibody (ET7110-55) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-55) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Villin1 was immunoprecipitated from 0.2 mg Caco-2 cell lysate with <a href="/products/ET7110-55" style="font-weight: bold;text-decoration: underline;">ET7110-55</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/ET7110-55" style="font-weight: bold;text-decoration: underline;">ET7110-55</a> at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: Caco-2 cell lysate (input)<br />Lane 2: <a href="/products/ET7110-55" style="font-weight: bold;text-decoration: underline;">ET7110-55</a> IP in Caco-2 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/ET7110-55" style="font-weight: bold;text-decoration: underline;">ET7110-55</a> in Caco-2 cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 6 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    Villin1 was immunoprecipitated from 0.2 mg Caco-2 cell lysate with ET7110-55 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET7110-55 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: Caco-2 cell lysate (input)
    Lane 2: ET7110-55 IP in Caco-2 cell lysate
    Lane 3: Rabbit IgG instead of ET7110-55 in Caco-2 cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 6 seconds; ECL: K1801

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Citation