Recombinant Anti-EpCAM antibody [EPR20532-222] (ab213500)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20532-222] to EpCAM
- Suitable for: IHC-P, WB, IP, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-EpCAM antibody [EPR20532-222]
See all EpCAM primary antibodies -
Description
Rabbit monoclonal [EPR20532-222] to EpCAM -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WB, IP, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: A431, T-47D, SK-OV-3, HT-29, HCT 116, MCF7 and 4T1 whole cell lysates; Human breast, colon and fetal kidney lysates; Mouse small intestine lysate. IHC-P: Human colon, breast carcinoma and endometrium cancer tissues; Mouse and rat colon tissues. ICC/IF: HCT 116 and 4T1 cells. IP: HCT 116 whole cell lysate.
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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. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20532-222 -
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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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab213500 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 |
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IHC-P |
1/16000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB | (1) |
1/1000. Detects a band of approximately 36, 40 kDa (predicted molecular weight: 35 kDa).
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IP |
1/40.
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ICC/IF |
1/100.
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Notes |
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IHC-P
1/16000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Detects a band of approximately 36, 40 kDa (predicted molecular weight: 35 kDa). |
IP
1/40. |
ICC/IF
1/100. |
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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Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling EpCAM with ab213500 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab214880) Ready to use.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab214880).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Positive staining on human breast carcinoma. The section was incubated with ab213500 at 4°C overnight. -
All lanes : Anti-EpCAM antibody [EPR20532-222] (ab213500) 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: 35 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 [EPR20532-222] 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, ab213500 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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All lanes : Anti-EpCAM antibody [EPR20532-222] (ab213500) at 1/1000 dilution
Lane 1 : Wild-type A431 whole cell lysate
Lane 2 : EPCAM knockout A431 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 35 kDa
Observed band size: 35 kDaLanes 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 A431 wild-type cells in Western blot. Loss of signal was observed when EpCAM knockout sample was used. A431 wild-type 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.
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Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling EpCAM with ab213500 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Membranous staining on rat colon is observed [PMID: 15637741].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 100% methanol-fixed HCT 116 (Human colorectal carcinoma cell line) cells labeling EpCAM with ab213500 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing membranous staining on HCT 116 cells.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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Immunohistochemical analysis of paraffin-embedded human colon tissue labeling EpCAM with ab213500 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Membranous staining on human colon is observed [PMID: 15637741].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-EpCAM antibody [EPR20532-222] (ab213500) at 1/2000 dilution
Lane 1 : T-47D (Human ductal breast epithelial tumor cell line) whole cell lysate
Lane 2 : SK-OV-3 (Human ovarian cancer cell line) whole cell lysate
Lane 3 : HT-29 (Human colorectal adenocarcinoma cell line) whole cell lysate
Lane 4 : HCT 116 (Human colorectal carcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 35 kDa
Observed band size: 36,40 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1: 15 seconds; Lane 2: 3 minutes; Lane 3: 10 seconds; Lane 4: 15 seconds.
The MW observed is consistent with the literature: PMID19136966; PMID 23618806.
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EpCAM was immunoprecipitated from 0.35 mg of HCT 116 (Human colorectal carcinoma cell line) whole cell lysate with ab213500 at 1/40 dilution.
Western blot was performed from the immunoprecipitate using ab213500 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.
Lane 1: HCT 116 whole cell lysate 10 µg (Input).
Lane 2: ab213500 IP in HCT 116 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab213500 in HCT 116 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
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Immunofluorescent analysis of 100% methanol-fixed 4T1 (Mouse mammary gland carcinoma cell line) cells labeling EpCAM with ab213500 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing membranous staining on 4T1 cells.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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Immunohistochemical analysis of paraffin-embedded human endometrium cancer tissue labeling EpCAM with ab213500 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Membranous staining on tumor cells of human endometrium cancer is observed [PMID: 15637741].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-EpCAM antibody [EPR20532-222] (ab213500) at 1/2000 dilution
Lane 1 : Human breast cancer lysate
Lane 2 : 4T1 (Mouse mammary gland carcinoma cell line) whole cell lysate
Lane 3 : Human colon lysate
Lane 4 : Mouse small intestine lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 35 kDa
Observed band size: 36,40 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1/2: 3 minutes; Lane 3: 10 seconds; Lane 4: 15 seconds.
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Anti-EpCAM antibody [EPR20532-222] (ab213500) at 1/1000 dilution + Human fetal kidney lysate at 10 µg
Secondary
VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution
Predicted band size: 35 kDa
Observed band size: 36,40 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling EpCAM with ab213500 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Membranous staining on mouse colon is observed [PMID: 15637741].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (14)
ab213500 has been referenced in 14 publications.
- Hu Y et al. Hepatoblastoma: Derived Exosomal LncRNA NEAT1 Induces BMSCs Differentiation into Tumor-Supporting Myofibroblasts via Modulating the miR-132/MMP9 Axis. J Oncol 2022:7630698 (2022). PubMed: 35300348
- Sun B et al. miR-2392 functions as tumour suppressor and inhibits malignant progression of hepatocellular carcinoma via directly targeting JAG2. Liver Int 42:1658-1673 (2022). PubMed: 35485355
- Khalaj K et al. Autophagy Is Impaired in Fetal Hypoplastic Lungs and Rescued by Administration of Amniotic Fluid Stem Cell Extracellular Vesicles. Am J Respir Crit Care Med 206:476-487 (2022). PubMed: 35671495
- Xie ZQ et al. Capsaicin suppresses hepatocarcinogenesis by inhibiting the stemness of hepatic progenitor cells via SIRT1/SOX2 signaling pathway. Cancer Med 11:4283-4296 (2022). PubMed: 35674129
- Lei X et al. Mitochondrial fission induces immunoescape in solid tumors through decreasing MHC-I surface expression. Nat Commun 13:3882 (2022). PubMed: 35794100