Recombinant Anti-EpCAM antibody [E144] (ab32392)
![Western blot - Anti-EpCAM antibody [E144] (ab32392)](https://www.abcam.com/ps/products/32/ab32392/Images/ab32392-484605-anti-epcam-antibody-e144-western-blot-wildtype-hct-116-epcam-knockout-hct-116-a431-mcf7.jpg "Western blot - Anti-EpCAM antibody [E144] (ab32392)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E144] to EpCAM
- Suitable for: WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-EpCAM antibody [E144]
See all EpCAM primary antibodies -
Description
Rabbit monoclonal [E144] to EpCAM -
Host species
Rabbit -
Tested applications
Suitable for: WBmore details
Unsuitable for: ICC/IF,IHC-Fr or IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HCT 116, A431, MCF7, Mouse colon tissue, and Rat colon tissue.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
E144 -
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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KO cell lines
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KO cell lysates
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Positive Controls
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32392 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 |
|---|---|---|
| WB | (3) |
1/1000 - 1/10000. Predicted molecular weight: 39 kDa.
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| Notes |
|---|
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WB
1/1000 - 1/10000. Predicted molecular weight: 39 kDa. |
Target
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Function
May act as a physical homophilic interaction molecule between intestinal epithelial cells (IECs) and intraepithelial lymphocytes (IELs) at the mucosal epithelium for providing immunological barrier as a first line of defense against mucosal infection. Plays a role in embryonic stem cells proliferation and differentiation. Up-regulates the expression of FABP5, MYC and cyclins A and E. -
Tissue specificity
Highly and selectively expressed by undifferentiated rather than differentiated embryonic stem cells (ESC). Levels rapidly diminish as soon as ESC's differentiate (at protein levels). Expressed in almost all epithelial cell membranes but not on mesodermal or neural cell membranes. Found on the surface of adenocarcinoma. -
Involvement in disease
Defects in EPCAM are the cause of diarrhea type 5 (DIAR5) [MIM:613217]. It is an intractable diarrhea of infancy characterized by villous atrophy and absence of inflammation, with intestinal epithelial cell dysplasia manifesting as focal epithelial tufts in the duodenum and jejunum.
Defects in EPCAM are a cause of hereditary non-polyposis colorectal cancer type 8 (HNPCC8) [MIM:613244]. HNPCC is a disease associated with marked increase in cancer susceptibility. It is characterized by a familial predisposition to early-onset colorectal carcinoma (CRC) and extra-colonic tumors of the gastrointestinal, urological and female reproductive tracts. HNPCC is reported to be the most common form of inherited colorectal cancer in the Western world. Clinically, HNPCC is often divided into two subgroups. Type I is characterized by hereditary predisposition to colorectal cancer, a young age of onset, and carcinoma observed in the proximal colon. Type II is characterized by increased risk for cancers in certain tissues such as the uterus, ovary, breast, stomach, small intestine, skin, and larynx in addition to the colon. Diagnosis of classical HNPCC is based on the Amsterdam criteria: 3 or more relatives affected by colorectal cancer, one a first degree relative of the other two; 2 or more generation affected; 1 or more colorectal cancers presenting before 50 years of age; exclusion of hereditary polyposis syndromes. The term 'suspected HNPCC' or 'incomplete HNPCC' can be used to describe families who do not or only partially fulfill the Amsterdam criteria, but in whom a genetic basis for colon cancer is strongly suspected. Note=HNPCC8 results from heterozygous deletion of 3-prime exons of EPCAM and intergenic regions directly upstream of MSH2, resulting in transcriptional read-through and epigenetic silencing of MSH2 in tissues expressing EPCAM. -
Sequence similarities
Belongs to the EPCAM family.
Contains 1 thyroglobulin type-1 domain. -
Post-translational
modificationsHyperglycosylated in carcinoma tissue as compared with autologous normal epithelia. Glycosylation at Asn-198 is crucial for protein stability. -
Cellular localization
Lateral cell membrane. Cell junction > tight junction. Co-localizes with CLDN7 at the lateral cell membrane and tight junction. - Information by UniProt
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Database links
- Entrez Gene: 4072 Human
- Entrez Gene: 17075 Mouse
- Entrez Gene: 171577 Rat
- Omim: 185535 Human
- SwissProt: P16422 Human
- SwissProt: Q99JW5 Mouse
- SwissProt: O55159 Rat
- Unigene: 542050 Human
see all -
Alternative names
- 17 1A antibody
- 323/A3 antibody
- Adenocarcinoma associated antigen antibody
see all
Images
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Western blot - Anti-EpCAM antibody [E144] (ab32392)All lanes : Anti-EpCAM antibody [E144] (ab32392) at 1/1000 dilution
Lane 1 : Wild-type HCT 116 cell lysate
Lane 2 : EPCAM knockout HCT 116 cell lysate
Lane 3 : A431 cell lysate
Lane 4 : MCF7 cell lysate
Lane 5 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 39 kDa
Observed band size: 37-45 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-EpCAM antibody [E144] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32392 was shown to bind specifically to EpCAM. A band was observed at 37/45 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in EPCAM knockout cell line ab281596 (knockout cell lysate ab282948). To generate this image, wild-type and EPCAM knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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![Western blot - Anti-EpCAM antibody [E144] (ab32392)](https://www.abcam.com/ps/products/32/ab32392/Images/ab32392-464583-AntiEpCAM-antibody-western-blot.jpg)
Western blot - Anti-EpCAM antibody [E144] (ab32392)All lanes : Anti-EpCAM antibody [E144] (ab32392) at 1/1000 dilution
Lane 1 : Mouse colon tissue lysate
Lane 2 : Rat small intestine tissue lysate
Lane 3 : Rat colon tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 39 kDa -
![Western blot - Anti-EpCAM antibody [E144] (ab32392)](https://www.abcam.com/ps/products/32/ab32392/Images/ab32392-412981-anti-epcam-antibody-e144-western-blot-a431-wt-a431-epcam-knockout-mcf7-hela2.jpg)
Western blot - Anti-EpCAM antibody [E144] (ab32392)All lanes : Anti-EpCAM antibody [E144] (ab32392) at 1/2000 dilution
Lane 1 : Wild-type A431 cell lysate
Lane 2 : EPCAM knockout A431 cell lysate
Lane 3 : MCF7 cell lysate
Lane 4 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 39 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?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 A431 cells in western blot with loss of signal observed in EpCAM knockout sample. Wild-type and EpCAM knockout A431 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.
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![Western blot - Anti-EpCAM antibody [E144] (ab32392)](https://www.abcam.com/ps/products/32/ab32392/Images/ab32392-44691-anti-epcam-antibody-e144-western-blot.jpg)
Western blot - Anti-EpCAM antibody [E144] (ab32392)Anti-EpCAM antibody [E144] (ab32392) at 1/2500 dilution + A431 cell lysate
Predicted band size: 39 kDa -
![Anti-EpCAM antibody [E144] (ab32392)](https://www.abcam.com/ps/products/32/ab32392/Images/ab32392-9-benefits-of-recombinant-antibodies.png)
Anti-EpCAM antibody [E144] (ab32392)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (55)
ab32392 has been referenced in 55 publications.
- Zhang PD et al. Feasibility of primary human cell cultures as a model for adamantinomatous craniopharyngioma research: Evidence from RNA-Seq analysis. Oncol Lett 19:2346-2354 (2020). PubMed: 32194734
- Jia N et al. The Human Lung Glycome Reveals Novel Glycan Ligands for Influenza A Virus. Sci Rep 10:5320 (2020). PubMed: 32210305
- Das B et al. Congenital Tufting Enteropathy-Associated Mutant of Epithelial Cell Adhesion Molecule Activates the Unfolded Protein Response in a Murine Model of the Disease. Cells 9:N/A (2020). PubMed: 32290509
- Karagonlar ZF et al. A Novel Function for KLF4 in Modulating the De-differentiation of EpCAM-/CD133- nonStem Cells into EpCAM+/CD133+ Liver Cancer Stem Cells in HCC Cell Line HuH7. Cells 9:N/A (2020). PubMed: 32408542
- Steins A et al. High-grade mesenchymal pancreatic ductal adenocarcinoma drives stromal deactivation through CSF-1. EMBO Rep 21:e48780 (2020). PubMed: 32173982