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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [4]
- Immunocytochemistry [1]
- Immunohistochemistry [2]
- Flow cytometry [2]
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- Product number
- MA5-15887 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PKN2 Monoclonal Antibody (1D1)
- Antibody type
- Monoclonal
- Antigen
- Purifed from natural sources
- Description
- MA5-15887 targets PRK2 in FACS, IHC, and WB applications and shows reactivity with Human, mouse, Non-human primate, and Rat samples.
- Antibody clone number
- 1D1
- Concentration
- Conc. Not Determined
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of PKN2 was achieved by transfecting HeLa cells with PKN2 specific validated siRNAs (Silencer® select Product #s11118, s11117). Western blot analysis (Fig. a) was performed using whole cell extracts from the PKN2 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with PKN2 Monoclonal Antibody (Product # MA5-15887, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 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 PKN2.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1), A-431 (Lane 2), K-562 (Lane 3) and HEK-293 (Lane 4). The blot was probed with Anti-PKN2 Monoclonal Antibody (Product # MA5-15887, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 130 kDa band corresponding to PKN2 was observed across the cell lines tested.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of PRK2 using a PRK2 monoclonal antibody (Product # MA5-15887) against a human PRK2 (AA: 555-718) recombinant protein.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of PRK2 using PRK2 monoclonal antibody (Product # MA5-15887) in PC-12 (1), COS-7 (2), K562 (3), Jurkat (4), HeLa (5), A431 (6), C6 (7), NIH/3T3 (8) and HEK293 (9) cell lysate.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of PKN2 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 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with PKN2 Monoclonal Antibody (1D1) (Product # MA5-15887) at 1:100 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) 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 and 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
- Immunohistochemical analysis of paraffin-embedded prostate tissues using PRK2 monoclonal antibody (Product # MA5-15887) followed with DAB staining.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded lung cancer tissues using PRK2 monoclonal antibody (Product # MA5-15887) followed with DAB staining.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometric analysis of NIH/3T3 cells using PRK2 monoclonal antibody (Product # MA5-15887) (blue) and negative control (red).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometric analysis of NIH/3T3 cells using PRK2 monoclonal antibody (Product # MA5-15887) (blue) and negative control (red).
