- MA5-17154 - Invitrogen Antibodies PPM1A antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot was performed using Anti-PPM1A Monoclonal Antibody (7F12) (Product # MA5-17154) and a 42 kDa band corresponding to Protein phosphatase 1A was observed across cell lines and tissues. (30 µg lysate) of MCF7 (Lane 1), THP-1 (Lane 2), HeLa (Lane 3),Rat Brain (Lane 4), Rat Kidney (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein 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 (1:1000) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000) 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: Western blot analysis of PPM1A using PPM1A monoclonal antibody (Product # MA5-17154) in Jurkat (1), Jurkat (2), A431 (3), HeLa (4), HEK293 (5), Raji (6), MCF-7 (7), and COS-7 (8) cell lysate.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of Protein phosphatase 1A was achieved by transfecting HeLa with Protein phosphatase 1A specific siRNAs (Silencer® select Product # S10919, S10921). Western Blot analysis (Fig. a) was performed using Whole cell extracts from the Protein phosphatase 1A knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The Blot was probed with PPM1A Monoclonal Antibody (7F12) (Product # MA5-17154, 1:1000 ) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000). Densitometric analysis of this western Blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Protein phosphatase 1A.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot was performed using Anti-PPM1A Monoclonal Antibody (7F12) (Product # MA5-17154) and a 42 kDa band corresponding to Protein phosphatase 1A was observed across cell lines and tissues. (30 µg lysate) of MCF7 (Lane 1), THP-1 (Lane 2), HeLa (Lane 3),Rat Brain (Lane 4), Rat Kidney (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein 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 (1:1000) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000) 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: Western blot analysis of PPM1A using a PPM1A monoclonal antibody (Product # MA5-17154) against a human PPM1A recombinant protein.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of PPM1A using PPM1A monoclonal antibody (Product # MA5-17154) in HEK293 (1) and PPM1A (AA: 202-382) human IgG Fc transfected HEK293 (2) cell lysate.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of PPM1A using PPM1A monoclonal antibody (Product # MA5-17154) in Jurkat (1), Jurkat (2), A431 (3), HeLa (4), HEK293 (5), Raji (6), MCF-7 (7), and COS-7 (8) cell lysate.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Protein phosphatase 1A 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 PPM1A Monoclonal Antibody (7F12) (Product # MA5-17154) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723), (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 Cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometric analysis of HeLa cells using PPM1A monoclonal antibody (Product # MA5-17154) (green) and negative control (red).

MA5-17154