PA1-841

antibody from Invitrogen Antibodies

Targeting: NRIP1 RIP140

Western blot

Immunoprecipitation

Antibody data

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Validation data

Product number: PA1-841 - Provider product page
Provider: Invitrogen Antibodies
Product name: RIP140 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: PA1-841 detects recombinant human Receptor-Interacting Protein 140 (RIP 140). PA1-841 has been successfully used in Western blot and immunoprecipitation procedures. By Western blot, this antibody detects two proteins at ~140 kDa representing human RIP 140 expressed in rabbit reticulocyte lysate. The PA1-841 immunogen is a synthetic peptide corresponding to residues G(718) N P T K G R V K K K E K T P L R D(735) of human RIP140. This immunizing peptide (Cat. # PEP-030) is available for use in neutralization and control experiments.
Reactivity: Human
Host: Rabbit
Isotype: IgG
Vial size: 100 µg
Concentration: 1 mg/mL
Storage: -20° C, Avoid Freeze/Thaw Cycles

NRIP1 protein structure - PA1-841 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of RIP140 was performed using 70% confluent log phase MCF7 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 RIP140 Polyclonal Antibody (Product # PA1-841) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731), (1:3000 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 nucleus localization. Mild non specific cytoplasmic staining was also observed. 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 RIP140 was achieved by transfecting MCF7 cells with RIP140 specific siRNA (Silencer® select Product # S15701, S15702). Immunofluorescence analysis was performed on untransfected MCF7 cells (panel a,d), transfected with non-specific scrambled siRNA (panels b,e) and transfected with RIP140 specific siRNA (panel c,f). Cells were fixed, permeabilized, and labelled with RIP140 Polyclonal Antibody (Product # PA1-841, 1:500 dilution) followed by Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731), (1:3000dilution). Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962), and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (Red) staining. Complete knockdown of RIP140 in siRNA transfected cells of specific signal was observed upon siRNA mediated knockdown (panel c,f) confirming specificity of the antibody to RIP140 (Green). The Images were captured at 60X magnification.