WB
Western blot analysis of PKA C-alpha on different lysates with Rabbit anti-PKA C-alpha antibody at 1/2,000 dilution. Lane 1: HEK-293 cell lysate Lane 2: PC-3 cell lysate Lane 3: MCF7 cell lysate Lane 4: PC-12 cell lysate Lane 5: C6 cell lysate Lane 6: COS-1 cell lysate Lysates/proteins at 15 µg/Lane. Predicted band size: 41 kDa Observed band size: 41 kDa Exposure time: 20 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-PKA C-alpha antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IF
Immunocytochemistry analysis of MCF7 cells labeling PKA C-alpha with Rabbit anti-PKA C-alpha antibody at 1/50 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-PKA C-alpha antibody at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of HEK-293 cells labeling PKA C-alpha. Cells were fixed and permeabilized. Then stained with the primary antibody (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
PKA C-alpha was immunoprecipitated from 0.2 mg HEK-293 cell lysate with at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HEK-293 cell lysate (input) Lane 2: IP in HEK-293 cell lysate Lane 3: Rabbit IgG instead of in HEK-293 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 3 minutes.| Product Name | PKA C-alpha Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within human PKA C-alpha aa 1-351. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCIFIHCIPWB |
| WB:1:2,000 IHC:1:200-1:1,000 IF:1:50 FC:1:1,000 IP: 1-2μg/sample |
|
| Species Reactivity | HumanMonkeyMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | PRKACA |
|---|---|
| Gene Synonyms | CAFD1 PKACA PPNAD4 |
| Gene Full Name | protein kinase cAMP-activated catalytic subunit alpha |
| Gene Summary | This gene encodes one of the catalytic subunits of protein kinase A, which exists as a tetrameric holoenzyme with two regulatory subunits and two catalytic subunits, in its inactive form. cAMP causes the dissociation of the inactive holoenzyme into a dimer of regulatory subunits bound to four cAMP and two free monomeric catalytic subunits. Four different regulatory subunits and three catalytic subunits have been identified in humans. cAMP-dependent phosphorylation of proteins by protein kinase A is important to many cellular processes, including differentiation, proliferation, and apoptosis. Constitutive activation of this gene caused either by somatic mutations, or genomic duplications of regions that include this gene, have been associated with hyperplasias and adenomas of the adrenal cortex and are linked to corticotropin-independent Cushing's syndrome. Alternative splicing results in multiple transcript variants encoding different isoforms. Tissue-specific isoforms that differ at the N-terminus have been described, and these isoforms may differ in the post-translational modifications that occur at the N-terminus of some isoforms. [provided by RefSeq, Jan 2015] |
| Alternative Names | cAMP-dependent protein kinase catalytic subunit alpha PKA C-alpha PRKACA PKACA |
| Molecular Weight(MW) | 41 kDa |
| Cellular Localization | Cell membrane,Cell projection,Cilium,Cytoplasm,Cytoplasmic vesicle |
WB
Western blot analysis of PKA C-alpha on different lysates with Rabbit anti-PKA C-alpha antibody at 1/2,000 dilution. Lane 1: HEK-293 cell lysate Lane 2: PC-3 cell lysate Lane 3: MCF7 cell lysate Lane 4: PC-12 cell lysate Lane 5: C6 cell lysate Lane 6: COS-1 cell lysate Lysates/proteins at 15 µg/Lane. Predicted band size: 41 kDa Observed band size: 41 kDa Exposure time: 20 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-PKA C-alpha antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IF
Immunocytochemistry analysis of MCF7 cells labeling PKA C-alpha with Rabbit anti-PKA C-alpha antibody at 1/50 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-PKA C-alpha antibody at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of HEK-293 cells labeling PKA C-alpha. Cells were fixed and permeabilized. Then stained with the primary antibody (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
PKA C-alpha was immunoprecipitated from 0.2 mg HEK-293 cell lysate with at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HEK-293 cell lysate (input) Lane 2: IP in HEK-293 cell lysate Lane 3: Rabbit IgG instead of in HEK-293 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 3 minutes.| Application Notes | WB:1:2,000 IHC:1:200-1:1,000 IF:1:50 FC:1:1,000 IP: 1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
