Rabbit Recombinant Monoclonal EPCAM antibody. Suitable for IP, WB, ICC/IF, IHC-P and reacts with Human, Mouse, Rat samples. Cited in 20 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
IP | WB | ICC/IF | IHC-P | |
---|---|---|---|---|
Human | Tested | Tested | Tested | Tested |
Mouse | Expected | Tested | Tested | Tested |
Rat | Expected | Expected | Expected | Tested |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/40 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/1000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/100 | Notes - |
Species Human | Dilution info 1/100 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/16000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Rat | Dilution info 1/16000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/16000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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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.
GA733-2, M1S2, M4S1, MIC18, TACSTD1, TROP1, M4S1, MIC18, TACSTD1, TROP1, EPCAM, GA733-2, M1S2, Epithelial cell adhesion molecule, Ep-CAM, Adenocarcinoma-associated antigen, Cell surface glycoprotein Trop-1, Epithelial cell surface antigen, Epithelial glycoprotein, Epithelial glycoprotein 314, KS 1/4 antigen, KSA, Major gastrointestinal tumor-associated protein GA733-2, Tumor-associated calcium signal transducer 1, EGP, EGP314, hEGP314
Rabbit Recombinant Monoclonal EPCAM antibody. Suitable for IP, WB, ICC/IF, IHC-P and reacts with Human, Mouse, Rat samples. Cited in 20 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EPR20532-222
Affinity purification Protein A
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
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.
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.
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.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
False colour image of Western blot: Anti-EpCAM antibody [EPR20532-222] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-EpCAM antibody [EPR20532-222] (ab213500) at 1/1000 dilution
Lane 1: Wild-type HCT 116 cell lysate at 20 µg
Lane 2: EPCAM knockout HCT 116 cell lysate at 20 µg
Lane 3: A431 cell lysate at 20 µg
Lane 4: MCF7 cell lysate at 20 µg
Lane 5: HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 35 kDa
Observed band size: 37-45 kDa
Lanes 1 - 3: Merged signal (red and green). Green - ab213500 observed at 40 kDa. Red - loading control, Anti-alpha Tubulin antibody [DM1A] - 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 Anti-alpha Tubulin antibody [DM1A] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed 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] (ab213500) 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 3: HeLa whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 35 kDa
Observed band size: 35 kDa
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) (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab213500 in HCT 116 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
All lanes: Immunoprecipitation - Anti-EpCAM antibody [EPR20532-222] (ab213500)
Predicted band size: 35 kDa
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) (Goat Anti-Rabbit IgG H&L (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 Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.
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) (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) (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.
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) (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) (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.
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.
All lanes: Western blot - Anti-EpCAM antibody [EPR20532-222] (ab213500) at 1/2000 dilution
Lane 1: T-47D (Human ductal breast epithelial tumor cell line) whole cell lysate at 20 µg
Lane 2: SK-OV-3 (Human ovarian cancer cell line) whole cell lysate at 20 µg
Lane 3: HT-29 (Human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 4: HCT 116 (Human colorectal carcinoma cell line) whole cell lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 35 kDa
Observed band size: 36 kDa, 40 kDa
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) (Goat Anti-Rabbit IgG H&L (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 Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.
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) (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) (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.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1/2: 3 minutes; Lane 3: 10 seconds; Lane 4: 15 seconds.
All lanes: Western blot - Anti-EpCAM antibody [EPR20532-222] (ab213500) at 1/2000 dilution
Lane 1: Human breast cancer lysate at 20 µg
Lane 2: 4T1 (Mouse mammary gland carcinoma cell line) whole cell lysate at 20 µg
Lane 3: Human colon lysate at 20 µg
Lane 4: Mouse small intestine lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 35 kDa
Observed band size: 36 kDa, 40 kDa
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-EpCAM antibody [EPR20532-222] (ab213500) at 1/1000 dilution
All lanes: Human fetal kidney lysate at 10 µg
All lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/4000 dilution
Predicted band size: 35 kDa
Observed band size: 36 kDa, 40 kDa
Exposure time: 3min
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) (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) (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.
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) (Goat Anti-Rabbit IgG H&L (HRP polymer) 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) (Goat Anti-Rabbit IgG H&L (HRP polymer) 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.
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