PKC alpha Recombinant Rabbit Monoclonal Antibody [SU31-08]
cat.: ET1608-15
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, IF-Tissue, IHC-P, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | SU31-08 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 77 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within Human PKC alpha aa 560-672 / 672. |
| Positive control: | 293T cell lysate, U-87 MG cell lysate, NIH/3T3 cell lysate, C6 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, Hela, MCF-7, CRC, A549, PC-12, 293T, NIH/3T3, C6. |
| Subcellular location: | Cytoplasm, Nucleus, Cell membrane, Mitochondrion membrane. |
| Recommended Dilutions:
WB IF-Cell IF-Tissue FC IP |
1:1,000-1:2,000 1:50-1:200 1:50-1:200 1:50-1:100 1:10-1:50 |
| Uniprot #: | SwissProt: P17252 Human | P20444 Mouse | P05696 Rat |
| Alternative names: | AAG6 Aging associated gene 6 aPKC KPCA_HUMAN PKC alpha PKC-A PKC-alpha PKCA PRKACA PRKCA Protein Kinase C alpha Protein kinase C alpha type |
Images
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Fig1:
Western blot analysis of PKC alpha on different lysates with Rabbit anti-PKC alpha antibody (ET1608-15) at 1/1,000 dilution. Lane 1: 293T cell lysate (20 µg/Lane) Lane 2: U-87 MG cell lysate (20 µg/Lane) Lane 3: NIH/3T3 cell lysate (20 µg/Lane) Lane 4: C6 cell lysate (20 µg/Lane) Lane 5: Mouse brain tissue lysate (40 µg/Lane) Lane 6: Rat brain tissue lysate (40 µg/Lane) Predicted band size: 77 kDa Observed band size: 77 kDa Exposure time: 9 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1608-15) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
PKC alpha was immunoprecipitated from 0.2 mg 293T cell lysate with ET1608-15 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1608-15 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: 293T cell lysate (input) Lane 2: ET1608-15 IP in 293T cell lysate Lane 3: Rabbit IgG instead of ET1608-15 in 293T cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 2 min; ECL: K1801 |
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Fig3:
Immunocytochemistry analysis of 293T cells labeling PKC alpha with Rabbit anti-PKC alpha antibody (ET1608-15) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PKC alpha antibody (ET1608-15) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig4:
Flow cytometric analysis of 293T cells labeling PKC alpha. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1608-15, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig5:
Flow cytometric analysis of NIH/3T3 cells labeling PKC alpha. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1608-15, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig6:
Flow cytometric analysis of C6 cells labeling PKC alpha. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1608-15, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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