Anti-EpCAM antibody [323/A3] (ab85987)
Key features and details
- Mouse monoclonal [323/A3] to EpCAM
- Suitable for: ICC/IF, Flow Cyt, WB
- Knockout validated
- Reacts with: Human
- Isotype: IgG1
Related conjugates and formulations
Overview
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Product name
Anti-EpCAM antibody [323/A3]
See all EpCAM primary antibodies -
Description
Mouse monoclonal [323/A3] to EpCAM -
Host species
Mouse -
Specificity
We have data to indicate that this antibody may not cross react with Rat. However, this has not been conclusively tested and expression levels may vary in certain cell lines/tissues. -
Tested applications
Suitable for: ICC/IF, Flow Cyt, WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
Tissue, cells or virus. MCF7 (human breast adenocarcinoma cell line) cells.
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Positive control
- Flow Cyt: A431 cells. ICC/IF: A431 and T47D cells. WB: MCF-7 cell lysate
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.40
Preservative: 0.097% Sodium azide
Constituent: PBS -
Concentration information loading...
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Purity
Protein A purified -
Purification notes
Purified from cell culture supernatant by protein A affinity chromatography, purity > 95% (by SDS-PAGE). -
Clonality
Monoclonal -
Clone number
323/A3 -
Isotype
IgG1 -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab85987 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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ICC/IF |
Use a concentration of 1 - 10 µg/ml.
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Flow Cyt |
Use a concentration of 1 - 4 µg/ml.
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WB |
Use a concentration of 1 - 2 µg/ml. Use under non reducing condition. Detects a band of approximately 40 kDa (predicted molecular weight: 35 kDa).
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Notes |
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ICC/IF
Use a concentration of 1 - 10 µg/ml. |
Flow Cyt
Use a concentration of 1 - 4 µg/ml. |
WB
Use a concentration of 1 - 2 µg/ml. Use under non reducing condition. Detects a band of approximately 40 kDa (predicted molecular weight: 35 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
- Omim: 185535 Human
- SwissProt: P16422 Human
- Unigene: 542050 Human
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Alternative names
- 17 1A antibody
- 323/A3 antibody
- Adenocarcinoma associated antigen antibody
see all
Images
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All lanes : Anti-EpCAM antibody [323/A3] (ab85987) at 2 µg/ml
Lane 1 : HEK293T/17 cell lysate (reducing)
Lane 2 : MCF7 cell lysate (reducing)
Lane 3 : HEK293T/17 cell lysate (non-reducing)
Lane 4 : MCF7 cell lysate (non-reducing)
Secondary
All lanes : IRDye800-conjugated anti-mouse IgG1
Predicted band size: 35 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted? -
Flow cytometry overlay histogram showing wild-type A431 (green line) and EPCAM knockout A431 cells stained with ab85987 (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 (ab85987) (1x106 in 100μl at 0.2 μg/ml) for 30 min at 4°C.
The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150117) was used at 1/2000 for 30 min at 4°C.
Isotype control antibody was mouse IgG1κ (ab170190) used at the same concentration and conditions as the primary antibody (wild-type A431 - black line; EPCAM knockout A431 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
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ab85987 staining EpCAM in T47D positive cells (top panel) and HeLa negative cells (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab85987 at 0.1μg/ml concentration and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (ab150080) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI. This antibody performed similarly using 100% methanol fixation. Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a single confocal section is shown.
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ab85987 staining EpCAM in wild-type A431 cells (top panel) and EpCAM knockout A431 cells (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab85987 at 0.1μg/ml concentration and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (ab150080) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI. This antibody performed similarly using 100% methanol fixation. Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a single confocal section is shown.
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ab85987 staining EpCAM in wild-type A431 cells (top panel) and EpCAM knockout A431 cells (bottom panel). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab85987 at 0.5μg/ml concentration and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (ab150080) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (10)
ab85987 has been referenced in 10 publications.
- Kapka-Skrzypczak L et al. IL-6 prevents CXCL8-induced stimulation of EpCAM expression in ovarian cancer cells. Mol Med Rep 19:2317-2322 (2019). PubMed: 30747214
- Lima LDP et al. Immunohistochemical Coexpression of Epithelial Cell Adhesion Molecule and Alpha-Fetoprotein in Hepatocellular Carcinoma. Can J Gastroenterol Hepatol 2018:5970852 (2018). PubMed: 30112355
- Jin X et al. Evaluation of Tumor-Derived Exosomal miRNA as Potential Diagnostic Biomarkers for Early-Stage Non-Small Cell Lung Cancer Using Next-Generation Sequencing. Clin Cancer Res 23:5311-5319 (2017). PubMed: 28606918
- Winter MJ et al. The epithelial cell adhesion molecule (Ep-CAM) as a morphoregulatory molecule is a tool in surgical pathology. Am J Pathol 163:2139-48 (2003). PubMed: 14633587
- Kirman I et al. Naturally occurring antibodies to epithelial cell adhesion molecule (EpCAM). Dig Dis Sci 48:2306-9 (2003). PubMed: 14714617
- Balzar M et al. Epidermal growth factor-like repeats mediate lateral and reciprocal interactions of Ep-CAM molecules in homophilic adhesions. Mol Cell Biol 21:2570-80 (2001). PubMed: 11259604
- Balzar M et al. Cytoplasmic tail regulates the intercellular adhesion function of the epithelial cell adhesion molecule. Mol Cell Biol 18:4833-43 (1998). PubMed: 9671492
- Litvinov SV et al. Ep-CAM: a human epithelial antigen is a homophilic cell-cell adhesion molecule. J Cell Biol 125:437-46 (1994). PubMed: 8163559
- Litvinov SV et al. Evidence for a role of the epithelial glycoprotein 40 (Ep-CAM) in epithelial cell-cell adhesion. Cell Adhes Commun 2:417-28 (1994). PubMed: 7842256
- Edwards DP et al. Monoclonal antibody identification and characterization of a Mr 43,000 membrane glycoprotein associated with human breast cancer. Cancer Res 46:1306-17 (1986). PubMed: 3510721