Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [2]
- Other assay [1]
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Validation data
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- Product number
- 703744 - Provider product page
- 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.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of PRMT3 was achieved by transfecting A549 cells with PRMT3 specific validated siRNA (Silencer® select Product # s19870). Western blot analysis (Fig a) was performed using whole cell extracts from PRMT3 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Anti-PRMT3 Recombinant Rabbit Monoclonal Antibody (Product # 703744) at a 1:5000 dilution and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036) (0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in the histogram (Fig b). Loss of signal upon siRNA mediated knockdown confirms that antibody is specific to PRMT3.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- 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).
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.
- 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.
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).