- MA1-16608 - Invitrogen Antibodies EP300 antibody

Submitted references HDAC1 modulates OGG1-initiated oxidative DNA damage repair in the aging brain and Alzheimer's disease.

Pao PC, Patnaik D, Watson LA, Gao F, Pan L, Wang J, Adaikkan C, Penney J, Cam HP, Huang WC, Pantano L, Lee A, Nott A, Phan TX, Gjoneska E, Elmsaouri S, Haggarty SJ, Tsai LH
Nature communications 2020 May 18;11(1):2484

Targeting of p300/CREB binding protein coactivators by simian virus 40 is mediated through p53.

Borger DR, DeCaprio JA
Journal of virology 2006 May;80(9):4292-303

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Detection of p300 in a HeLa nuclear extract using Product # MA1-16608 (1:250). ECL: 15 second exposure.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of p300 in HeLa nuclear extract. Samples were incubated in p300 monoclonal antibody (Product # MA1-16608 using a dilution of 1:250. ECL: 15 sec exposure.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of EP300 was performed with 10 µg of HeLa cells transfected with Transfection Reagent alone (Lane 1), 100nM Non-Targeting control siRNA (Lane 2), or 100nM siRNA against EP300 (Lane 3). Proteins were resolved using a NuPAGE® Novex 4-12% Bis-Tris Gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002), and a protein size ladder. Proteins were wet transferred to a Pierce Nitrocellulose Membrane (Product # 88025) OR Pierce PVDF Membrane (Product # 88518) and blocked with Pierce Starting Block T20 (PBS) Blocking Buffer (Product # 37539) for 1 hour at room temperature. EP300 was detected at ~ 300 kDa using EP300 Mouse monoclonal antibody (Product # MA1-16608) diluted in Pierce Starting Block T20 (PBS) Blocking Buffer 4°C overnight on a rocking platform. Pierce Goat Anti-Mouse (Product # 31437) HRP-Conjugated Antibodies at a 1:2500 dilution were used and chemiluminescent detection was performed using Pierce Supersignal West Dura Maximum Sensitivity Substrate (Product # 37071). Relative density of the bands normalized to Lamin A/C (74/65 kDa). EP300 Antibody (Product # MA1-16608) confirms silencing of EP300 expression.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of p300 Monoclonal Antibody (RW128) 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 45 minutes at room temperature. The cells were labeled with p300 Monoclonal Antibody (RW128) (Product # MA1-16608) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (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 dilution). Panel d represents the merged image showing nuclear as well as cytoplasmic 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: Knockdown of p300 Monoclonal Antibody (RW128) was achieved by transfecting HeLa cells with p300 specific siRNA (Silencer® select Product # S4696, S4697). Immunofluorescence analysis was performed on untransfected HeLa cells (panel a,d), transfected with non-specific scrambled siRNA (panels b,e) and transfected with p300 specific siRNA (panel c,f). Cells were fixed, permeabilized, and labelled with p300 Monoclonal Antibody (RW128) (Product # MA1-16608, 1:100 dilution) followed by Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2000 dilution). Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962), and Rhodamine Phalloidin (Product # R415, 1:300 dilution) was used for cytoskeletal F-actin (Red) staining. reduction in signal of specific signal was observed upon siRNA mediated knockdown (panel c,f) confirming specificity of the antibody to p300 (Green). The Images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry of p300 in THP-1 cells. Samples were incubated in p300 monoclonal antibody (Product # MA1-16608) and a matched isotype control using a dilution of 1 µg/mL for 30 minutes at room temperature followed by mouse F(ab)2 IgG (H+L) APC-conjugated secondary antibody. Antibody (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and permeabilized with 0.1% Saponin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry of p300 in Raw264.7 cells. Samples were incubated with p300 monoclonal antibody (Product # MA1-16608) using a dilution of 1.0 µg/mL for 30 minutes at room temperature followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (Product # 35503). Antibody (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin.

MA1-16608

Antibody data