- MA5-34812 - Invitrogen Antibodies BRD2 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of BRD2 in SiHa cell lysate. Samples were incubated with BRD2 monoclonal antibody (Product # MA5-34812), at a dilution of 1:2000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-BRD2 Recombinant Rabbit Monoclonal Antibody (JG40-42) (Product # MA5-34812) and a 110 kDa band corresponding to BRD2 was observed across cell lines tested. Nuclear enriched extracts (30 µg lysate) of HeLa (Lane 1), HEK-293 (Lane 2), MCF7 (Lane 3), A549 (Lane 4), SiHa (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:20,000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of BRD2 was achieved by transfecting HeLa with BRD2 specific siRNAs (Silencer® select Product # S12070, S12071). Western blot analysis (Fig. a) was performed using Nuclear enriched extracts from the BRD2 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2), and untransfected cells (lane 1). The blot was probed with BRD2 Recombinant Rabbit Monoclonal Antibody (JG40-42) (Product # MA5-34812, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000). Densitometric analysis of this western blot is shown in the histogram (Fig. b). A decrease in signal upon siRNA mediated knockdown confirms that the antibody is specific to BRD2.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of BRD2 was performed using 70% confluent log phase BeWo 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 45 minutes at room temperature. The cells were labeled with BRD2 Recombinant Rabbit Monoclonal Antibody (JG40-42) (Product # MA5-34812) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). 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 showing nuclear localization. Panel e represents control cells with no primary antibody to assess the background. The images were captured at 40X magnification in CellInsight CX7 LZR High-Content Screening (HCS) Platform (Product # CX7A1110LZR) and externally deconvoluted (D.Sage et al./Methods 115 (2017) 28–41).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of BRD2 in SiHa cells (green). Samples were fixed in paraformaldehyde and permeabilised with 0.25% Triton X100/PBS, incubated with BRD2 monoclonal antibody (Product # MA5-34812), followed by DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of BRD2 in A431 cells (green). Samples were fixed in paraformaldehyde and permeabilised with 0.25% Triton X100/PBS, incubated with BRD2 monoclonal antibody (Product # MA5-34812), followed by DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of BRD2 in 293T cells (green). Samples were fixed in paraformaldehyde and permeabilised with 0.25% Triton X100/PBS, incubated with BRD2 monoclonal antibody (Product # MA5-34812), followed by DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry analysis of BRD2 in paraffin-embedded human tonsil tissue. Samples were incubated with BRD2 monoclonal antibody (Product # MA5-34812), and followed by hematoxylin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry analysis of BRD2 in paraffin-embedded human colon tissue. Samples were incubated with BRD2 monoclonal antibody (Product # MA5-34812), and followed by hematoxylin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometry of BRD2 in 293T cells (purple) compared with an unlabelled control (cells without incubation with primary antibody; yellow). Samples were incubated with BRD2 monoclonal antibody (Product # MA5-34812) at a dilution of 1:100, followed by Alexa Fluor 488-conjugated goat anti-rabbit IgG.

MA5-34812