Explore
Validate
Learn
Western blotAntibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [7]
- Immunocytochemistry [1]
- Flow cytometry [1]
Submit
Validation data
Reference
Comment
Report error
- Product number
- MA5-17154 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PPM1A Monoclonal Antibody (7F12)
- Antibody type
- Monoclonal
- Antigen
- Purifed from natural sources
- Description
- MA5-17154 targets PPM1A in FACS, pep-ELISA, and WB applications and shows reactivity with Human and Non-human primate samples.
- Antibody clone number
- 7F12
- Concentration
- 1 mg/mL
No comments: Submit comment
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).
- 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.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- 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.
- 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).
- 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.
- 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.
- 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)
- Main image
- Experimental details
- Flow cytometric analysis of HeLa cells using PPM1A monoclonal antibody (Product # MA5-17154) (green) and negative control (red).
