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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Other assay [1]
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- Product number
- PA5-78129 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PKC zeta Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- Positive Control: 293T, A431, HeLa, HepG2
- Concentration
- 1 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of PKC zeta in whole cell lysate using 30 µg of protein. Samples were separated with 10% SDS-PAGE and incubated with PKC zeta polyclonal antibody (Product # PA5-78129) using a dilution of 1:1000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- PKC zeta Polyclonal Antibody detects PKC zeta protein by western blot analysis. Various whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with PKC zeta Polyclonal Antibody (Product # PA5-78129) diluted by 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-PKC zeta Polyclonal Antibody (Product # PA5-78129) and a 67kDa band corresponding to PKC zeta was observed in all cell lines and tissue lysates tested except HL-60 and Mouse Skeletal Muscle which are reported to be negative. Whole cell extracts (30 µg lysate) of HEK-293 (Lane 1), HeLa (Lane 2), Hep G2 (Lane 3), MDA-MB-231 (Lane 4), THP-1 (Lane 5), HL-60 (Lane 6), Mouse Brain (Lane 7) and Mouse Skeletal Muscle (Lane 8) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris 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 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) 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
- Immunofluorescence analysis of PKC zeta was performed using 70% confluent log phase HeLa and HL-60 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 1 hour at room temperature. The cells were labeled with PKC zeta Polyclonal Antibody (Product # PA5-78129) at 1:100 dilution 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) 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 endosome pattern of staining. Panel e shows HL-60 cells with no expression of PKC zeta. Panel f 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
- Immunoprecipitation of PKC zeta was performed in 293T whole cell extracts using 5 µg of PKC zeta Polyclonal Antibody (Product # PA5-78129). Samples were transferred to a membrane and probed with PKC zeta Polyclonal Antibody as a primary antibody and an HRP-conjugated anti-Rabbit IgG was used as a secondary antibody.
