- MA5-24291 - Invitrogen Antibodies EOMES antibody

Lin MY, Wang YL, Wu WL, Wolseley V, Tsai MT, Radic V, Thornton ME, Grubbs BH, Chow RH, Huang IC
Scientific reports 2017 Nov 1;7(1):14883

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis from lysates of BG01V human embryonic stem cells untreated (-) or mesendoderm differentiated (+). PVDF membrane was probed with 1 µg/mL of mouse Anti-human EOMES Monoclonal Antibody (Product # MA5-24291) followed by HRP-conjugated Anti-mouse IgG Secondary Antibody. Specific bands were detected for EOMES at approximately 100 and 80 kDa (as indicated). This experiment was conducted under reducing conditions.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of EOMES in BG01V human embryonic stem cells untreated (-) or mesendoderm differentiated (+). Samples were incubated in EOMES monoclonal antibody (Product # MA5-24291) using a dilution of 1 µg/mL followed by a HRP-conjugated Anti-Mouse IgG secondary antibody. Specific bands were detected for EOMES at approximately 100 and 80 kDa (as indicated). This experiment was conducted under reducing conditions.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-EOMES Monoclonal Antibody (644730) (Product # MA5-24291) and 75, 85kDa bands corresponding to EOMES were observed in NK-92 but were absent in A-431 and THP-1 which are reported to be negative. Modified whole cell extracts (1% SDS) (30 µg lysate) of NK-92 (Lane 1), A-431 (Lane 2) and THP-1 (Lane 3) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris 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 (1µg/mL) and 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). An uncharacterized band (*) was observed at ~60kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry analysis of EOMES in immersion fixed BG01V human embryonic stem cells differentiated into mesoderm. Samples were incubated in EOMES monoclonal antibody (Product # MA5-24291) using a dilution of 10 µg/mL for 3 hours at room temperature followed by NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red, upper panel) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometric analysis of BG01V human embryonic stem cells differentiated to mesendoderm were stained with mouse Anti-human EOMES Monoclonal Antibody (Product # MA5-24291) or isotype control antibodyopen histogram), followed by Allophycocyanin-conjugated Anti-mouse IgG Secondary Antibody. To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry of EOMES in BG01V human embryonic stem cells differentiated to mesendoderm. Samples were incubated in EOMES monoclonal antibody (Product # MA5-24291) or isotype control antibody followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody. To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Supportive validation

MA5-24291