- MA5-24297 - Invitrogen Antibodies RCOR1 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis from lysates of Jurkat human acute T cell leukemia cell line and MOLT-4 human acute lymphoblastic leukemia cell line. PVDF Membrane was probed with 1 µg/mL of human RCOR1/CoREST Monoclonal Antibody (Product # MA5-24297) followed by HRP-conjugated Anti-mouse IgG Secondary Antibody. A specific band was detected for RCOR1/CoREST at approximately 64-66 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 RCOR1 in Jurkat human acute T cell leukemia cell line and MOLT‚4 human acute lymphoblastic leukemia cell line. Samples were incubated in RCOR1 monoclonal antibody (Product # MA5-24297) using a dilution of 1 µg/mL followed by a HRP-conjugated Anti-Mouse IgG secondary antibody. A specific band was detected for RCOR1/CoREST at approximately 64-66 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-RCOR1 Monoclonal Antibody (658526), (Product # MA5-24297) and a 60 kDa band corresponding to RCOR1 was observed in the cell lines tested. Modified whole cell extracts (1%SDS) (30 µg lysate) of Jurkat (Lane 1), HeLa (Lane 2), HEL 92.1.7 (Lane 3) and K-562 (Lane 4) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX). 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 ug/ml) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP conjugate (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).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry analysis of RCOR1 in immersion fixed HeLa human cervical epithelial carcinoma cell line. Samples were incubated in RCOR1 monoclonal antibody (Product # MA5-24297) 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)
Main image
Experimental details: Immunofluorescence analysis of RCOR1 was performed using 70% confluent log phase 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 RCOR1 Monoclonal Antibody (658526) (Product # MA5-24297) at 5 µg/mL concentration in 0.1% BSA, incubated at 4 degree Celsius overnight and then with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant 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 represents the merged image showing nuclear localization. Panel e 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: Immunocytochemical analysis of RCOR1/CoREST was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using human RCOR1/CoREST Monoclonal Antibody (Product # MA5-24297) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the 557-conjugated Anti-mouse IgG Secondary Antibody (red, upper pane and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei.

MA5-24297