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Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit

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Catalog# HAK21033

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit

Application

  • WB

Reactivity

  • Human

General Overview

Kit Components

Product Includes Specification Application Reactivity Mw
Vimentin[ET1610-39] 20µl WB,IF-Cell,IF-Tissue,IHC-P,IP,FC,IHC-Fr Human,Mouse,Rat Predicted band size: 54 kDa
N Cadherin[ET1607-37] 20µl WB,IF-Tissue,IHC-P,FC,IHC-Fr Human,Mouse,Rat Predicted band size: 100 kDa
Claudin 1[HA721999] 20µl WB,IHC-P Human,Mouse,Rat Predicted band size: 23 kDa
Beta Catenin[ET1601-5] 20µl WB,IHC-P,IF-Tissue,IP,mIHC,IF-Cell,IHC-Fr,FC Human,Mouse,Rat Predicted band size: 85 kDa
SNAIL[ER1706-22] 20µl WB,IF-Cell,IHC-P Human,Mouse 29 kDa, additional band 35 kDa
SLUG[HA722828] 20µl WB,IF-Cell Human Predicted band size: 30 kDa
ZEB[HA721438] 20µl WB,IHC-P,FC Human,Mouse,Rat Predicted band size: 124 kDa
E-Cadherin[ET1607-75] 20µl WB,IF-Cell,IF-Tissue,IHC-P,FC,IP,mIHC Human Predicted band size: 97 kDa
ZO1[HA722797] 20µl WB,IF-Cell,IHC-P Human,Mouse,Rat,Monkey Predicted band size: 195 kDa
Goat Anti-Rabbit IgG (H+L)[HA1001] 100µl WB,ELISA,IHC-P Rabbit

Product Description

The Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit provides an economical means of evaluating EMT. The kit contains enough primary antibody to perform two western blots per primary.

Product Features

Storage Buffer

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

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Background

Epithelial-mesenchymal transition (EMT) is an essential process during development whereby epithelial cells aquire mesenchymal, fibroblast-like properties and display reduced intracellular adhesion and increased motility.EMT depends on a reduction in expression of cell adhesion molecules. </br>Cadherins mediate calcium-dependent cell-cell adhesion and play critical roles in normal tissue development. E-cadherin is considered an active suppressor of invasion and growth of many epithelial cancers . Recent studies indicate that cancer cells have up-regulated N-cadherin in addition to loss of E-cadherin. Zona occludens proteins ZO-1, 2, and 3 are peripheral membrane adaptor proteins that link junctional transmembrane proteins such as occludin and claudin to the actin cytoskeleton. ZO-1 and -2 are required for tight junction formation and function ; mutations in ZO-1 and Claudin induce EMT. </br>Vimentin's dynamic structural changes and spatial re-organization in response to extracellular stimuli helps to coordinate various signaling pathways . β-catenin is a key downstream effector in the Wnt signaling pathway .β-catenin also activates Slug. Slug (SNAI2) is a widely expressed transcriptional repressor and member of the Snail family of zinc finger transcription factors . The binding of Slug to integrin promoter sequences represses integrin expression and results in reduced cell adhesion . One of the targets suppressed by ZEB proteins is E-cadherin.

Data Links

Background References

1. Aigner K, Dampier B, Descovich L, Mikula M, Sultan A, Schreiber M, Mikulits W, Brabletz T, Strand D, Obrist P, Sommergruber W, Schweifer N, Wernitznig A, Beug H, Foisner R, Eger A. The transcription factor ZEB1 (deltaEF1) promotes tumour cell dedifferentiation by repressing master regulators of epithelial polarity. Oncogene. 2007 Oct 25;26(49):6979-88.

2. Moreno-Bueno G, Portillo F, Cano A. Transcriptional regulation of cell polarity in EMT and cancer. Oncogene. 2008 Nov 24;27(55):6958-69.

3. Manfioletti G, Fedele M. Epithelial-Mesenchymal Transition (EMT) 2021. Int J Mol Sci. 2022 May 23;23(10):5848.

Images

  • Western blot analysis of Vimentin on different lysates with Rabbit anti-Vimentin antibody (<a href="/products/ET1610-39" style="font-weight: bold;text-decoration: underline;">ET1610-39</a>) at 1/20,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (10 µg/Lane)<br />Lane 2: HEK-293 cell lysate (10 µg/Lane)<br />Lane 3: Jurkat cell lysate (10 µg/Lane)<br />Lane 4: C2C12 cell lysate (10 µg/Lane)<br />Lane 5: RAW264.7 cell lysate (10 µg/Lane)<br />Lane 6: C6 cell lysate (10 µg/Lane)<br /><br />Predicted band size: 54 kDa<br />Observed band size: 54 kDa<br />Exposure time: 14 seconds;<br />4-20% SDS-PAGE gel.<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/ET1610-39" style="font-weight: bold;text-decoration: underline;">ET1610-39</a>) at 1/20,000 dilution and competitor's antibody at 1/1,000 dilution were 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.

    Western blot analysis of Vimentin on different lysates with Rabbit anti-Vimentin antibody (ET1610-39) at 1/20,000 dilution and competitor's antibody at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (10 µg/Lane)
    Lane 2: HEK-293 cell lysate (10 µg/Lane)
    Lane 3: Jurkat cell lysate (10 µg/Lane)
    Lane 4: C2C12 cell lysate (10 µg/Lane)
    Lane 5: RAW264.7 cell lysate (10 µg/Lane)
    Lane 6: C6 cell lysate (10 µg/Lane)

    Predicted band size: 54 kDa
    Observed band size: 54 kDa
    Exposure time: 14 seconds;
    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 (ET1610-39) at 1/20,000 dilution and competitor's antibody at 1/1,000 dilution were 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.

  • Western blot analysis of N Cadherin on different lysates with Rabbit anti-N Cadherin antibody (<a href="/products/ET1607-37" style="font-weight: bold;text-decoration: underline;">ET1607-37</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />Lane 1: 293T cell lysate<br />Lane 2: A549 cell lysate<br />Lane 3: HeLa cell lysate<br />Lane 4: A-172 cell lysate<br />Lane 5: MCF7 cell lysate (negative)<br />Lane 6: C2C12 cell lysate<br />Lane 7: C6 cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br />Predicted band size: 100 kDa<br />Observed band size: 140-150 kDa<br />Exposure time: 2 minutes 6 seconds;<br />4-20% SDS-PAGE gel.<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/ET1607-37" style="font-weight: bold;text-decoration: underline;">ET1607-37</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were 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.

    Western blot analysis of N Cadherin on different lysates with Rabbit anti-N Cadherin antibody (ET1607-37) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.

    Lane 1: 293T cell lysate
    Lane 2: A549 cell lysate
    Lane 3: HeLa cell lysate
    Lane 4: A-172 cell lysate
    Lane 5: MCF7 cell lysate (negative)
    Lane 6: C2C12 cell lysate
    Lane 7: C6 cell lysate

    Lysates/proteins at 15 µg/Lane.
    Predicted band size: 100 kDa
    Observed band size: 140-150 kDa
    Exposure time: 2 minutes 6 seconds;
    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 (ET1607-37) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were 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.

  • Western blot analysis of E-Cadherin on different lysates with Rabbit anti-E-Cadherin antibody (<a href="/products/ET1607-75" style="font-weight: bold;text-decoration: underline;">ET1607-75</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />Lane 1: MCF7 cell lysate<br />Lane 2: MDA-MB-231 cell lysate (negative)<br />Lane 3: HT-29 cell lysate<br />Lane 4: HCT 116 cell lysate<br />Lane 5: A431 cell lysate<br />Lane 6: Caco-2 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br />Predicted band size: 97 kDa<br />Observed band size: 80~120 kDa<br />Exposure time: 43 seconds;<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/ET1607-75" style="font-weight: bold;text-decoration: underline;">ET1607-75</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were 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.

    Western blot analysis of E-Cadherin on different lysates with Rabbit anti-E-Cadherin antibody (ET1607-75) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.

    Lane 1: MCF7 cell lysate
    Lane 2: MDA-MB-231 cell lysate (negative)
    Lane 3: HT-29 cell lysate
    Lane 4: HCT 116 cell lysate
    Lane 5: A431 cell lysate
    Lane 6: Caco-2 cell lysate

    Lysates/proteins at 20 µg/Lane.
    Predicted band size: 97 kDa
    Observed band size: 80~120 kDa
    Exposure time: 43 seconds;
    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 (ET1607-75) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were 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.

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