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
- PA5-30127 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PPP2R3A Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: NT2D1, U87-MG, MCF-7.
- Concentration
- 1 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of A-431 (Lane 1), U-87 MG (Lane 2) and HCT 116 (Lane 3). The blot was probed with Anti-PPP2R3A Polyclonal Antibody (Product # PA5-30127, 1:500 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 150 kDa band corresponding to PPP2R3A was observed across the cell lines tested.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using PPP2R3A Polyclonal Antibody (Product # PA5-30127). Sample (30 µg of whole cell lysate). Lane A: NT2D1. Lane B: U87-MG. 5% SDS PAGE. PPP2R3A Polyclonal Antibody (Product # PA5-30127) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using PPP2R3A Polyclonal Antibody (Product # PA5-30127). Various tissue extracts (50 µg) were separated by 5% SDS-PAGE, and the membrane was blotted with PPP2R3A Polyclonal Antibody (Product # PA5-30127) diluted at 1:500. 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 PPP2R3A was achieved by transfecting U-87 MG cells with PPP2R3A specific siRNA (Silencer® select Product # s10977). Western blot analysis (Fig. a) was performed using whole cell extracts from the PPP2R3A knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with PPP2R3A Polyclonal Antibody (Product # PA5-30127, 1:500 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 histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to PPP2R3A.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of A-431 (Lane 1), U-87 MG (Lane 2) and HCT 116 (Lane 3). The blot was probed with Anti-PPP2R3A Polyclonal Antibody (Product # PA5-30127, 1:500 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 150 kDa band corresponding to PPP2R3A was observed across the cell lines tested.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- PPP2R3A Polyclonal Antibody detects PPP2R3A protein at cytoplasm by immunofluorescent analysis. Sample: U87-MG cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: PPP2R3A protein stained by PPP2R3A Polyclonal Antibody (Product # PA5-30127) diluted at 1:500. Blue: Hoechst 33342 staining.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of PPP2R3A was performed using 70% confluent log phase U-87 MG cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with PPP2R3A Polyclonal Antibody (Product # PA5-30127) at 5µg/mL in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). 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 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using PPP2R3A (Product # PA5-30127) antibody at 1:250 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.