MA5-12604
antibody from Invitrogen Antibodies
Targeting: EPCAM
17-1A, 323/A3, CD326, CO-17A, EGP-2, EGP34, EGP40, Ep-CAM, ESA, GA733-2, HEA125, KS1/4, KSA, Ly74, M4S1, MH99, MIC18, MK-1, MOC31, TACST-1, TACSTD1, TROP1
Antibody data
- Antibody Data
- Antigen structure
- References [4]
- Comments [0]
- Validations
- Western blot [1]
- Immunocytochemistry [2]
- Immunohistochemistry [3]
- Flow cytometry [1]
- Other assay [1]
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Validation data
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- Product number
- MA5-12604 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- EpCAM Monoclonal Antibody (Ber-EP4)
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- MA5-12604 targets Epithelial Specific Antigen in IHC (P) applications and shows reactivity with Human samples. The MA5-12604 immunogen is mCF-7 human breast carcinoma cell line.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- Ber-EP4
- Vial size
- 500 µL
- Concentration
- Conc. Not Determined
- Storage
- 4° C
Submitted references Quercetin Regulates Key Components of the Cellular Microenvironment during Early Hepatocarcinogenesis.
Identification of cells with colony-forming activity, self-renewal capacity, and multipotency in ovarian endometriosis.
Primary localized malignant biphasic mesothelioma of the liver in a patient with asbestosis.
Primary localized malignant biphasic mesothelioma of the liver in a patient with asbestosis.
Reyes-Avendaño I, Reyes-Jiménez E, González-García K, Pérez-Figueroa DC, Baltiérrez-Hoyos R, Tapia-Pastrana G, Sánchez-Chino XM, Villa-Treviño S, Arellanes-Robledo J, Vásquez-Garzón VR
Antioxidants (Basel, Switzerland) 2022 Feb 11;11(2)
Antioxidants (Basel, Switzerland) 2022 Feb 11;11(2)
Identification of cells with colony-forming activity, self-renewal capacity, and multipotency in ovarian endometriosis.
Chan RW, Ng EH, Yeung WS
The American journal of pathology 2011 Jun;178(6):2832-44
The American journal of pathology 2011 Jun;178(6):2832-44
Primary localized malignant biphasic mesothelioma of the liver in a patient with asbestosis.
Sasaki M, Araki I, Yasui T, Kinoshita M, Itatsu K, Nojima T, Nakanuma Y
World journal of gastroenterology 2009 Feb 7;15(5):615-21
World journal of gastroenterology 2009 Feb 7;15(5):615-21
Primary localized malignant biphasic mesothelioma of the liver in a patient with asbestosis.
Sasaki M, Araki I, Yasui T, Kinoshita M, Itatsu K, Nojima T, Nakanuma Y
World journal of gastroenterology 2009 Feb 7;15(5):615-21
World journal of gastroenterology 2009 Feb 7;15(5):615-21
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockout of EpCAM was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, AssayID CRISPR701274_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Immunofluorescence analysis was performed on wild type A-431 cells (panel a,d), A-431 Cas9 control cells (panel b,e), and A-431 EpCAM KO cells (panel c, f). Cells were fixed, permeabilized, and labeled with EpCAM Monoclonal Antibody (Ber-EP4) (Product # MA5-126048203, 1:100 dilution), followed by Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ Plus 488 (Product # A32723, 1:2000). Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962), and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Loss of signal (panel c,f) upon CRISPR mediated knockout (KO) confirms that the antibody is specific to EpCAM (green). The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of EpCAM was done on 70% confluent log phase A431 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with of EpCAM (Ber-EP4) Mouse Monoclonal Antibody (Product # MA5-12604) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing membranous localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of EpCAM was performed using HT-29 and BJ cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with EpCAM Mouse Monoclonal Antibody (Product # MA5-12604) at 1:250 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green) in HT-29 cells. Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image of HT-29 cells, which is a positive model for EpCAM expression showing a plasma membrane localization. Panel e represents the merged image of BJ cells, that are null for EpCAM protein expression. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Formalin-fixed, paraffin-embedded human breast cancer stained with Epithelial Specific Antigen antibody using peroxidase-conjugate and AEC chromogen. Note membrane staining of tumor cells.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of EpCAM/CD326 showing staining in the membrane of paraffin-embedded human colon carcinoma tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a EpCAM/CD326 Mouse Monoclonal Antibody (Ber-EP4) (Product # MA5-12604) diluted in 3% BSA-PBS at a dilution of 1:50 for 1 hour at 37°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of EpCAM/CD326 showing staining in the cytoplasm and nucleus of paraffin-embedded human prostrate carcinoma tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a EpCAM/CD326 Mouse Monoclonal Antibody (Ber-EP4) (Product # MA5-12604) diluted in 3% BSA-PBS at a dilution of 1:50 for 1 hour at 37°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry analysis of EpCAM / CD326 was done on HT-29 cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with EpCAM / CD326 Mouse Monoclonal Antibody (MA512604, red histogram) or with mouse isotype control (yellow histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Rabbit Anti-Mouse Secondary Antibody (A11059) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 7 Effect of quercetin on EpCAM-positive CSCs population. ( a ) Representative images of immunohistochemical staining for EpCAM in both control (NT, NT + Q) and MRHM (CT and CT + Q) liver tissues. Positive area is shown in brown gradients. Photomicrographs were taken at 10x and 40x magnification. ( b ) Quantification of positive area for EpCAM in CT and CT + Q groups. Bars represent the mean +- SD, p < 0.001 (***). EpCAM: epithelial cell adhesion molecule, NT: untreated, NT + Q: untreated plus quercetin, CT: carcinogenic treatment, CT + Q: carcinogenic treatment plus quercetin.