- MA1-23275 - Invitrogen Antibodies RBBP4 antibody

Qian YW, Lee EY
The Journal of biological chemistry 1995 Oct 27;270(43):25507-13

Qian YW, Lee EY
The Journal of biological chemistry 1995 Oct 27;270(43):25507-13

Qian YW, Wang YC, Hollingsworth RE Jr, Jones D, Ling N, Lee EY
Nature 1993 Aug 12;364(6438):648-52

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using RbAp48 Monoclonal Antibody (13D10) (Product # MA1-23275). SH-SY5Y whole cell and nuclear extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with RbAp48 Monoclonal Antibody (13D10) (Product # MA1-23275) diluted at 1:2,000. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of RbAp48 was performed by separating 30 µg of SH-SY-5Y whole cell and nuclear extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a RbAp48 Monoclonal Antibody (13D10) (Product # MA1-23275) at a dilution of 1:2000. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of RbAp48 was achieved by transfecting HeLa with RbAp48 specific siRNAs (Silencer® select Products # s11838, s11837). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from the RbAp48 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with RbAp48 Monoclonal Antibody (13D10) (Product # MA1-23275, 1 µg/mL) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Reduction in signal upon siRNA mediated knock down confirms that antibody is specific to RbAp48.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-RbAp48 Monoclonal Antibody (13D10) (Product # MA1-23275) and a 50 kDa band corresponding to RbAp48 was observed in all cell and tissue lysates tested. Modified whole cell lysates (1% SDS) (30 µg lysate) of HeLa (Lane 1), A-431 (Lane 2), Hep G2 (Lane 3), PANC-1 (Lane 4), tissue lysates (30 µg lysate) of Mouse Spleen (Lane 5), Rat Lung (Lane 6) and Rat Spleen (Lane 7) were electrophoresed using NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX). 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 µg/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).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry-Immunofluorescence analysis of RbAp48 was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: RbAp48 Monoclonal Antibody (13D10) (Product # MA1-23275) diluted at 1:200. Red: phalloidin, a cytoskeleton marker. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of RbAp48 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 RbAp48 Monoclonal Antibody (13D10) (Product # MA1-23275) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 (Product # A28175) at a dilution of 1:2000 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 background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: MA1-23275 antibody immunoprecipitates RbAp48 protein-DNA in ChIP experiments. ChIP Sample: HeLa whole cell lysate/extract with 5 µg preimmune mouse IgG (A) or 5 µg of RbAp48 antibody (Product # MA1-23275) (B). The precipitated DNA was detected by PCR with primer set targeting to SOX9 gene locus.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunoprecipitation of RbAp48 was performed in HeLa whole cell extract using 5 µg of RbAp48 Monoclonal Antibody (13D10) (Product # MA1-23275). Samples were transferred to a membrane and probed with RbAp48 Monoclonal Antibody (13D10) as a primary antibody and an HRP-conjugated HRP-conjugated anti mouse IgG was used as a secondary antibody.

MA1-23275