703744

antibody from Invitrogen Antibodies

Targeting: PRMT3 HRMT1L3

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Data presented on provider website)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

703744

Provider

Invitrogen Antibodies

Product name

PRMT3 Recombinant Rabbit Monoclonal Antibody (23H2L13)

Antibody type

Monoclonal

Antigen

Other

Reactivity

Human, Rat

Host

Rabbit

Isotype

IgG

Antibody clone number

23H2L13

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunocytochemistry analysis, A549 cells were fixed and permeabilized for detection of endogenous PRMT3 using Anti-PRMT3 Recombinant Rabbit Monoclonal Antibody (Product # 703744) at a 1:100 dilution and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a 1:2000 dilution. Panel a) shows representative cells that were stained for detection and localization of PRMT3 protein (green), Panel b) is stained for nuclei (blue) using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). Panel c) represents cytoskeletal F-actin staining using Rhodamine Phalloidin (Product # R415) at a 1:300 dilution. Panel d) is a composite image of Panels a, b and c clearly demonstrating cytoplasmic localization of PRMT3. 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 PRMT3 was achieved by transfecting A549 cells with specific validated siRNA (Silencer® select Product # s19870). Immunofluorescence analysis was performed on A549 cells (untransfected, panel a-d), transfected with PRMT3 specific siRNA (panel i-l) or non-specific scrambled siRNA (panels e-h). Cells were fixed, permeabilized, and labeled with Anti-PRMT3 Recombinant Rabbit Monoclonal Antibody (Product # 703744) at a 1:100 dilution, followed by Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a 1:2000 dilution. Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962), and Rhodamine Phalloidin (Product # R415) at a 1:300 dilution was used for cytoskeletal F-actin (red) staining. Significant reduction of the signal was observed upon siRNA mediated knockdown (panel i-l) confirming the specificity of the antibody to PRMT3 (green). The images were captured at 60X magnification.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

PRMT3 was immunoprecipitated using 5 µg of the Anti-PRMT3 Recombinant Rabbit Monoclonal Antibody (Product # 703744) from whole cell extracts (800µg) of A549 (Lane 3) using the Protein A/G Dynabeads® (Product # 10001D and Product # 10003D). Normal Rabbit IgG was used as a Isotype control (Lane 2). Western blot analysis was performed using Anti-PRMT3 Recombinant Rabbit Monoclonal Antibody (Product # 703744) at a 1:5000 dilution. The blot was detected by chemiluminescence using Peroxidase IgG Fraction Monoclonal Mouse Anti-Rabbit IgG, light chain specific antibody (Product # 211-032-171) at a 1:10000 dilution. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10% Bis-Tris gel (Product # NP03011BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).