Invitrogen

PRMT3 Recombinant Rabbit Monoclonal Antibody (23H2L13)

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated.

View all (15) PRMT3 antibodies

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Datasheet

Protocols

Questions & Answers

Tech Support

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PRMT3 Antibody in Immunocytochemistry (ICC/IF) — 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.

PRMT3 Antibody in Western Blot (WB) — Western blot was performed using Anti-PRMT3 Recombinant Rabbit Monoclonal Antibody (Product # 703744) and a 60 kDa band corresponding to PRMT3 was observed across the cell lines tested. Whole cell extracts (30 µg lysate) of A549 (Lane 1), HeLa (Lane 2), MDA-MB-231 (Lane 3), MCF7 (Lane 4), T-47D (Lane 5), U-2 OS (Lane 6), LNCaP (Lane 7) and PC-12 (Lane 8) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). 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 (1:5000 dilution) and detected by chemiluminescence Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036) at a 1:4000 dilution. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

PRMT3 Antibody in Immunoprecipitation (IP) — 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).

PRMT3 Antibody — Antibody specificity was demonstrated by siRNA-mediated knockdown of the target protein. A549 cells were transfected with PRMT3 specific siRNA and decrease in signal intensity was observed in immunofluorescence application using Anti-PRMT3 Recombinant Rabbit Monoclonal Antibody (Product # 703744). {KD}

Product Details

703744

Applications	Tested Dilution	Publications
Western Blot (WB)	1:5,000	-
Immunocytochemistry (ICC/IF)	1:100	-
Immunoprecipitation (IP)	2.5 µg/million cells	-

Product Specifications
Species Reactivity	Human, Rat
Host/Isotype	Rabbit / IgG
Class	Recombinant Monoclonal
Type	Antibody
Clone	23H2L13
Immunogen	Protein corresponding to human PRMT3 [aa331-aa531] View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.5 mg/mL
Purification	Protein A
Storage buffer	PBS, pH 7.4
Contains	0.09% sodium azide
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2815338

Product Specific Information

This antibody is predicted to react with Mouse, Bovine.

Target Information

Type I protein arginine N-methyltransferases (PRMTs), such as PRMT3, catalyze the formation of asymmetric N(G),N(G)-dimethylarginine (ADMA) residues in proteins.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: heterogeneous nuclear ribonucleoprotein methyltransferase-like 3; Heterogeneous nuclear ribonucleoprotein methyltransferase-like protein 3; HMT1 hnRNP methyltransferase-like 3; Protein arginine N-methyltransferase 3; protein arginine N-methyltransferase 3(hnRNP methyltransferase S. cerevisiae)-like 3

Gene Aliases: HRMT1L3; PRMT3

UniProt ID: (Human) O60678, (Rat) O70467

Entrez Gene ID: (Human) 10196, (Rat) 89820

Molecular Function: protein modifying enzyme