JavaScript is disabled in your browser. Please enable JavaScript to view this website.
AB261708

Human EPCAM knockout A-431 cell lysate

Be the first to review this product! Submit a review

|

(0 Publication)

EPCAM KO cell lysate available now. KO validated by Immunocytochemistry, Next Generation Sequencing, Western blot. Free of charge wild type control included. Knockout achieved by CRISPR/Cas9 X = 2 bp deletion Frameshift: 99.58%.

View Alternative Names

17 1A, 323/A3, AUA1, Adenocarcinoma-associated antigen, Antigen identified by monoclonal antibody AUA1, CD326, CD326 antigen, CO 17A, CO17 1A, Cell surface glycoprotein Trop-1, Cell surface glycoprotein Trop-2, DIAR5, EGP 2, EGP314, EGP40, EPCAM_HUMAN, ESA, EpCAM1, Epithelial Cell Adhesion Molecule Intracellular Domain (EpCAM-ICD), Epithelial cell adhesion molecule, Epithelial cell surface antigen, Epithelial cellular adhesion molecule, Epithelial glycoprotein, Epithelial glycoprotein 1, Epithelial glycoprotein 314, GA733 1, GA733-2, GP4, HNPCC8, Human epithelial glycoprotein 2, KS 1/4 antigen, KS1/4, KSA, Ly74, Lymphocyte antigen 74, M1S 1, M1S2, M4S1, MIC18, MK 1, Major gastrointestinal tumor-associated protein GA733-2, Membrane component chromosome 4 surface marker (35kD glycoprotein), Membrane component, chromosome 4, surface marker, Membrane component, chromosome 4, surface marker 1, Protein 289A, TACD1, TACSTD1, TROP1, Tumor associated calcium signal transducer 2 precursor, Tumor-associated calcium signal transducer 1, gastrointestinal tumor-associated antigen 2, 35-KD glycoprotein, hEGP 2, hEGP314, mEGP314

5 Images
Western blot - Human EPCAM knockout A-431 cell lysate (AB261708)
  • WB

Lab

Western blot - Human EPCAM knockout A-431 cell lysate (AB261708)

Lane 1 : Wild-type A431 cell lysate 20 ug

Lane 2 : EPCAM knockout A431 cell lysate 20 ug

Lane 3 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate 20 ug

Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug

Lanes 1 - 4 : Merged signal (red and green). Green - ab32392 observed at 40 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

ab32392 was shown to react with EpCAM in wild-type A-431 cells in western blot with loss of signal observed in EpCAM knockout cell line ab261902 (knockout cell lysate ab263942). Wild-type and EpCAM knockout A-431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab32392 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-EpCAM antibody [E144] (<a href='/en-us/products/primary-antibodies/epcam-antibody-e144-ab32392'>ab32392</a>) at 1/2000 dilution

Lane 1:

Wild-type A431 cell lysate at 20 µg

Lane 2:

EPCAM knockout A431 cell lysate at 20 µg

Lane 2:

Western blot - Human EPCAM knockout A-431 cell line (<a href='/en-us/products/cell-lines/human-epcam-knockout-a-431-cell-line-ab261902'>ab261902</a>)

Lane 3:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 4:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Predicted band size: 35 kDa

Observed band size: 40 kDa

false

Western blot - Human EPCAM knockout A-431 cell lysate (AB261708)
  • WB

Lab

Western blot - Human EPCAM knockout A-431 cell lysate (AB261708)

Lane 1 : Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate 20 ug

Lane 2 : EPCAM knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate 20 ug

Lane 3 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate 20 ug

Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug

Lanes 1 - 4 : Merged signal (red and green). Green - ab224826 observed at 35 kDa. Red - loading control, ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.

ab224826 was shown to react with EpCAM in wild-type A-431 cells in Western blot Loss of signal was observed when EpCAM knockout cell line ab261902 (knockout cell lysate ab263942) was used. Wild-type A-431 and EpCAM knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab224826 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at a 1 in 400 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-EpCAM antibody [4A7] - C-terminal (<a href='/en-us/products/primary-antibodies/epcam-antibody-4a7-c-terminal-ab224826'>ab224826</a>) at 1/400 dilution

Lane 1:

Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

EPCAM knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human EPCAM knockout A-431 cell line (<a href='/en-us/products/cell-lines/human-epcam-knockout-a-431-cell-line-ab261902'>ab261902</a>)

Lane 3:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 4:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Predicted band size: 35 kDa

Observed band size: 35 kDa

false

Western blot - Human EPCAM knockout A-431 cell lysate (AB261708)
  • WB

Lab

Western blot - Human EPCAM knockout A-431 cell lysate (AB261708)

Lane 1 : Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate

Lane 2 : EPCAM knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate

Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate

Lanes 1 - 3 : Merged signal (red and green). Green - ab213500 observed at 40 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

ab213500 was shown to react with EpCAM in wild-type A-431 cells in Western blot Loss of signal was observed when EpCAM knockout cell line ab261902 (knockout cell lysate ab263942) was used. Wild-type A-431 and EpCAM knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab213500 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-EpCAM antibody [EPR20532-222] (<a href='/en-us/products/primary-antibodies/epcam-antibody-epr20532-222-ab213500'>ab213500</a>) at 1/1000 dilution

Lane 1:

Wild-type A431 whole cell lysate at 20 µg

Lane 2:

EPCAM knockout A431 whole cell lysate at 20 µg

Lane 2:

Western blot - Human EPCAM knockout A-431 cell line (<a href='/en-us/products/cell-lines/human-epcam-knockout-a-431-cell-line-ab261902'>ab261902</a>)

Lane 3:

HeLa whole cell lysate at 20 µg

Predicted band size: 35 kDa

Observed band size: 35 kDa

false

Western blot - Human EPCAM knockout A-431 cell lysate (AB261708)
  • WB

Lab

Western blot - Human EPCAM knockout A-431 cell lysate (AB261708)

Lane 1 : Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate

Lane 2 : EPCAM knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate

Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate

Lanes 1 - 3 : Merged signal (red and green). Green - ab223582 observed at 40 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

ab223582 was shown to react with EpCAM in wild-type A-431 cells in Western blot Loss of signal was observed when EpCAM knockout cell line ab261902 (knockout cell lysate ab263942) was used. Wild-type A-431 and EpCAM knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab223582 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-EpCAM antibody [EPR20532-225] (<a href='/en-us/products/primary-antibodies/epcam-antibody-epr20532-225-ab223582'>ab223582</a>) at 1/1000 dilution

Lane 1:

Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

EPCAM knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human EPCAM knockout A-431 cell line (<a href='/en-us/products/cell-lines/human-epcam-knockout-a-431-cell-line-ab261902'>ab261902</a>)

Lane 3:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Predicted band size: 35 kDa

Observed band size: 40 kDa

false

Next Generation Sequencing - Human EPCAM knockout A-431 cell lysate (AB261708)
  • NGS

Supplier Data

Next Generation Sequencing - Human EPCAM knockout A-431 cell lysate (AB261708)

Knockout achieved by CRISPR/Cas9; X = 2 bp deletion; Frameshift : 99.58%

Key facts

Cell type

A-431

Species or organism

Human

Tissue

Skin

Knockout validation

Immunocytochemistry,Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9 X = 2 bp deletion Frameshift: 99.58%

Disease

Epidermoid Carcinoma

style
left-column

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
style
left-column

Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

style
left-column

What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab261708-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human EPCAM knockout A431 cell lysate", "number":"AB261708-CMP02", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type A-431 cell lysate", "number":"AB261708-CMP01", "productcode":"" } ] } } }
style
left-column

Properties and storage information

Gene name
EPCAM
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Immunocytochemistry, Next Generation Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
style
left-column

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

EpCAM also known as epithelial cell adhesion molecule is a transmembrane glycoprotein with a molecular weight of around 40 kDa. Alternative names include CD326 and TACSTD1. This protein is expressed on the surface of most epithelial cells and plays a role in cell adhesion. It is involved in maintaining the integrity of epithelial tissues by mediating homotypic cell-cell interactions which are important for the structure and function of tissues.
Biological function summary

EpCAM facilitates signaling pathways that impact cellular processes like proliferation and differentiation. It is not a part of a larger protein complex but interacts directly with other cellular machinery to transmit intracellular signals. EpCAM influences stem cell behavior and is important in tissue homeostasis assisting in both normal regenerative processes and wound healing.

Pathways

EpCAM participates in regulatory mechanisms involving the Wnt/β-catenin signaling pathway which affects gene expression and cell fate decisions. Through its action EpCAM modulates components like E-cadherin which is important for cell adhesion and the maintenance of the epithelial layer. It indirectly influences pathways relating to MYC a transcription factor that regulates cell cycle progression and apoptosis.

EpCAM is associated with various types of cancer including colorectal and breast cancer. Its overexpression often correlates with aggressive cancer phenotypes and poor prognosis. Besides cancers EpCAM mutations can lead to congenital tufting enteropathy a rare condition affecting intestinal epithelial structure and function. Aberrant EpCAM expression affects β-catenin levels impacting cellular pathways linked to tumor progression and epithelial integrity.
style
left-column

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

style
left-column

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Female

style
left-column

Product protocols

style
left-column
style
left-column
style
right-column

Complementary Products

Primary Antibodies

AB237385

Alexa Fluor® 647 Anti-EpCAM antibody [EPR20533-63]

RabMAb|Recombinant

Flow Cytometry - Alexa Fluor® 647 Anti-EpCAM antibody [EPR20533-63] (AB237385)

  • 0
  • 0
Assay Kits

AB242247

Mouse Epcam ELISA Kit

Recombinant|SimpleStep

Sandwich ELISA - Mouse Epcam ELISA Kit (AB242247)

  • 0
  • 0
Proteins & Peptides

AB151338

Recombinant Human EpCAM protein

  • 0
  • 0

Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com