Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [5]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
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Validation data
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- Product number
- PA5-27643 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PPP2R1A Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: HeLa, NIH-3T3, JC, BCL-1.
- Concentration
- 0.12 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot analysis of PPP2R1A using 30 µg of HeLa lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a PPP2R1A polyclonal antibody (Product # PA5-27643) at a dilution of 1:1000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of PPP2R1A was performed by separating 30 µg of Whole cell extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a PPP2R1A Polyclonal Antibody (Product # PA5-27643) at a dilution of 1:1000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using PPP2R1A Polyclonal Antibody (Product # PA5-27643). Sample (30 µg of whole cell lysate). Lane A: NIH-3T3. Lane B: JC. Lane C: BCL-1. 7.5% SDS PAGE. PPP2R1A Polyclonal Antibody (Product # PA5-27643) diluted at 1:5,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of PPP2R1A was achieved by transfecting HeLa with PPP2R1A specific siRNAs (Silencer® select Product # S10964, S10965). Western blot analysis (Fig. a) was performed using Whole cell extracts from the PPP2R1A knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with PPP2R1A Polyclonal Antibody (Product # PA5-27643, 1:1000 ) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 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 PPP2R1A.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-PPP2R1A Polyclonal Antibody(Product # PA5-27643) and a 60kDa band corresponding to PPP2R1A was observed across all the tested cell lines and tissues. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), MDA-MB-231 (Lane 2), MCF7 (Lane 3), Jurkat (Lane 4), NIH/3T3 (Lane 5), L6 (Lane 6), Mouse Brain (Lane 7), Rat Brain (Lane 8) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). 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-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 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
- Immunofluorescent analysis of PPP2R1A in methanol-fixed HeLa cells using a PPP2R1A polyclonal antibody (Product # PA5-27643) at a 1:500 dilution.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of PPP2R1A was performed using 70% confluent log phase HeLa 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 PPP2R1A Polyclonal Antibody (Product # PA5-27643) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (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 a predominant nuclear localization. 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
- Immunohistochemistry (Paraffin) analysis of PPP2R1A was performed in paraffin-embedded mouse kidney tissue using PPP2R1A Polyclonal Antibody (Product # PA5-27643) at a dilution of 1:2500.