Rabbit Recombinant Monoclonal EPCAM antibody. Suitable for IP, Flow Cyt, WB, ICC/IF, IHC-P and reacts with Mouse samples. Cited in 7 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
IP | Flow Cyt | WB | ICC/IF | IHC-P | |
---|---|---|---|---|---|
Mouse | Tested | Tested | Tested | Tested | Tested |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/30 | 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 | Dilution info 1/500 | Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/5000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/100 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/2000 | 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 (By similarity).
Tacstd1, Epcam, Tacstd1, Epithelial cell adhesion molecule, Ep-CAM, Epithelial glycoprotein 314, Protein 289A, Tumor-associated calcium signal transducer 1, EGP314, mEGP314
Rabbit Recombinant Monoclonal EPCAM antibody. Suitable for IP, Flow Cyt, WB, ICC/IF, IHC-P and reacts with Mouse samples. Cited in 7 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EPR20533-63
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.
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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.
Flow cytometry overlay histogram showing left M158 positive cells and right negative NIH3T3 cells stained with ab221552 (red line). The cells were incubated in 1x PBS containing 10 % normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab221552) (1x106 in 100 µL at 0.1 µg/ml) for 30 min on ice./p>
The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor ® 488, pre-adsorbed) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) was used at 1/2000 for 30 min on ice.
Isotype control antibody (black line) was Rabbit IgG (monoclonal) (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.
Flow cytometric analysis of 4% paraformaldehyde-fixed 4T1 (mouse mammary gland carcinoma cell line) and NIH/3T3 (mouse embyro fibroblast cell line) cells labeling EpCAM with ab221552 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
Flow cytometry was performed with fresh cells without fixation and permeabilization.
Negative control: NIH/3T3 (PMID:23264216).
EpCAM was immunoprecipitated from 0.35 mg of mouse kidney lysate with ab221552 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab221552 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.
Lane 1: Mouse kidney lysate 10 μg (Input).
Lane 2: ab221552 IP in mouse kidney lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab221552 in mouse kidney lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds.
All lanes: Immunoprecipitation - Anti-EpCAM antibody [EPR20533-63] (ab221552)
Predicted band size: 35 kDa
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1/2: 3 seconds; Lane 3: 10 seconds.
The MW observed is consistent with the literature (PMID 23409978; PMID 23618806).
All lanes: Western blot - Anti-EpCAM antibody [EPR20533-63] (ab221552) at 1/5000 dilution
Lane 1: Mouse colon lysate at 20 µg
Lane 2: Mouse kidney lysate at 20 µg
Lane 3: 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
Developed using the ECL technique.
Predicted band size: 35 kDa
Observed band size: 39 kDa
Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling EpCAM with ab221552 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. 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) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized 4T1 (mouse mammary gland carcinoma cell line) and NIH/3T3 (mouse embyro fibroblast cell line) cells labeling EpCAM with ab221552 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (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.
Negative control: NIH/3T3 (PMID:23264216).
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.
EpCAM was immunoprecipitated from 0.35 mg of 4T1 (mouse mammary gland carcinoma cell line) whole cell lysate with ab221552 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab221552 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.
Lane 1: 4T1 (Mouse mammary gland carcinoma) whole cell lysate 10 μg (Input).
Lane 2: ab221552 IP in 4T1 whole cell lysate .
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab221552 in 4T1 whole cell lysate .
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
The band between 12-15kDa has been documented in literature as the cleaved form of EpCAM (PMID: 23409978).
All lanes: Immunoprecipitation - Anti-EpCAM antibody [EPR20533-63] (ab221552)
Predicted band size: 35 kDa
Blocking/Dilution buffer: 5% NFDM/TBST.
The MW observed is consistent with the literature (PMID 23409978; PMID 23618806).
Negative control: NIH/3T3 (PMID:23264216).
All lanes: Western blot - Anti-EpCAM antibody [EPR20533-63] (ab221552) at 1/5000 dilution
Lane 1: 4T1 (mouse mammary gland carcinoma cell line) whole cell lysate at 20 µg
Lane 2: NIH/3T3 (mouse embyro fibroblast 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
Developed using the ECL technique.
Predicted band size: 35 kDa
Observed band size: 39 kDa
Exposure time: 1s
Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling EpCAM with ab221552 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous staining on mouse lung 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) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Blocking/Dilution buffer: 5% NFDM/TBST.
The MW observed is consistent with the literature (PMID 23618806).
The band between 12-15kDa has been documented in literature as the cleaved form of EpCAM (PMID: 23409978).
Negative control: NIH/3T3 (PMID:23264216).
All lanes: Western blot - Anti-EpCAM antibody [EPR20533-63] (ab221552) at 1/5000 dilution
Lane 1: 4T1 (mouse mammary gland carcinoma cell line) whole cell lysate at 20 µg
Lane 2: NIH/3T3 (mouse embyro fibroblast 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
Developed using the ECL technique.
Predicted band size: 35 kDa
Observed band size: 12 kDa, 39 kDa
Exposure time: 10s
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