MA5-15640

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

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-15640

Provider

Invitrogen Antibodies

Product name

EpCAM Monoclonal Antibody (7E11)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Description

MA5-15640 targets EPCAM in IHC and WB applications and shows reactivity with Human samples.

Antibody clone number

7E11

Concentration

Conc. Not Determined

References

Submitted references

Epstein-Barr virus-associated lymphoepithelioma-like cholangiocarcinoma: a rare variant of intrahepatic cholangiocarcinoma with favourable outcome.

Chan AW, Tong JH, Sung MY, Lai PB, To KF
Histopathology 2014 Nov;65(5):674-83

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Expression of stemness markers (CD133 and EpCAM) in prognostication of hepatocellular carcinoma.

Chan AW, Tong JH, Chan SL, Lai PB, To KF
Histopathology 2014 Jun;64(7):935-50

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of CD326/EpCAM/Epithelial Specific antigen using CD326/EpCAM/Epithelial Specific antigen monoclonal antibody (Product # MA5-15640) in HTC116 (1) and T47D (2) cell lysate.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of CD326/EpCAM/Epithelial Specific antigen using CD326/EpCAM/Epithelial Specific antigen monoclonal antibody (Product # MA5-15640) in HTC116 (1) and T47D (2) cell lysate.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of CD326/EpCAM/Epithelial Specific antigen using CD326/EpCAM/Epithelial Specific antigen monoclonal antibody (Product # MA5-15640) in HTC116 (1) and T47D (2) cell lysate.

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, Assay ID CRISPR701274_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of EpCAM was performed by loading 30 µg of A-431 wild type (Lane 1), A-431 Cas9 (Lane 2) andA-431 EpCAM KO (Lane 3) membrane enriched extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-EpCAM Monoclonal Antibody (7E11) (Product # MA5-15640, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:5000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to EpCAM.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-EpCAM Mouse Monoclonal Antibody (Product # MA5-15640) and a 35 kDa band corresponding to EpCAM was observed in cell lines tested except for HeLa, Jurkat and BJ. Membrane enriched extracts (30 µg lysate) of HeLa (Lane 1), HCT 116 (Lane 2), HT-29 (Lane 3), Jurkat (Lane 4) and BJ (Lane 5) were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by detected by chemiluminescence with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of EpCAM was performed using HT-29 and HeLa 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 # MA5-15640) at 1:100 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723) 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 HeLa 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.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of paraffin-embedded human lung cancer (A), colon cancer (B), breast cancer (C) and rectal cancer (D), using CD326/EpCAM/Epithelial Specific antigen monoclonal antibody (Product # MA5-15640) followed with DAB staining.