Recombinant Anti-cAMP Protein Kinase Catalytic subunit antibody [EP2102Y] (ab76238)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP2102Y] to cAMP Protein Kinase Catalytic subunit
- Suitable for: ICC/IF, WB, IP, IHC-P, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-cAMP Protein Kinase Catalytic subunit antibody [EP2102Y]
See all cAMP Protein Kinase Catalytic subunit primary antibodies -
Description
Rabbit monoclonal [EP2102Y] to cAMP Protein Kinase Catalytic subunit -
Host species
Rabbit -
Specificity
The immunogen used for this product shares 92% homology with PKA C-beta and PKA C-gamma. Cross-reactivity with these proteins have not been confirmed experimentally.
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Tested applications
Suitable for: ICC/IF, WB, IP, IHC-P, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- ICC/IF: HeLa cells Flow Cyt (intra): MCF-7 and HeLa cells IHC-P: Rat stomach tissue, Mouse cerebrum tissue, Human testis and thyroid carcinoma Tissue WB: MCF-7, HeLa, NIH/3T3 and C6 cell lysates
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP2102Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab76238 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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ICC/IF |
1/100 - 1/250.
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WB |
1/20000 - 1/200000. Detects a band of approximately 42 kDa (predicted molecular weight: 46 kDa).
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IP |
1/40 - 1/50.
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IHC-P |
1/750. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/100 - 1/250. Use of HRP-conjugated or polymerized HRP secondary antibodies recommended, stronger signals have been found using the polymerized HRP secondary. |
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Flow Cyt (Intra) |
1/60 - 1/80.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Notes |
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ICC/IF
1/100 - 1/250. |
WB
1/20000 - 1/200000. Detects a band of approximately 42 kDa (predicted molecular weight: 46 kDa). |
IP
1/40 - 1/50. |
IHC-P
1/750. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/100 - 1/250. Use of HRP-conjugated or polymerized HRP secondary antibodies recommended, stronger signals have been found using the polymerized HRP secondary. |
Flow Cyt (Intra)
1/60 - 1/80. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Target
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Function
Phosphorylates a large number of substrates in the cytoplasm and the nucleus. Regulates the abundance of compartmentalized pools of its regulatory subunits through phosphorylation of PJA2 which binds and ubiquitinates these subunits, leading to their subsequent proteolysis. Phosphorylates CDC25B, ABL1, NFKB1, CLDN3, PSMC5/RPT6, PJA2, RYR2, RORA and VASP. RORA is activated by phosphorylation. Required for glucose-mediated adipogenic differentiation increase and osteogenic differentiation inhibition from osteoblasts. Involved in the regulation of platelets in response to thrombin and collagen; maintains circulating platelets in a resting state by phosphorylating proteins in numerous platelet inhibitory pathways when in complex with NF-kappa-B (NFKB1 and NFKB2) and I-kappa-B-alpha (NFKBIA), but thrombin and collagen disrupt these complexes and free active PRKACA stimulates platelets and leads to platelet aggregation by phosphorylating VASP. Prevents the antiproliferative and anti-invasive effects of alpha-difluoromethylornithine in breast cancer cells when activated. RYR2 channel activity is potentiated by phosphorylation in presence of luminal Ca(2+), leading to reduced amplitude and increased frequency of store overload-induced Ca(2+) release (SOICR) characterized by an increased rate of Ca(2+) release and propagation velocity of spontaneous Ca(2+) waves, despite reduced wave amplitude and resting cytosolic Ca(2+). PSMC5/RPT6 activation by phosphorylation stimulates proteasome. Negatively regulates tight junctions (TJs) in ovarian cancer cells via CLDN3 phosphorylation. NFKB1 phosphorylation promotes NF-kappa-B p50-p50 DNA binding. Involved in embryonic development by down-regulating the Hedgehog (Hh) signaling pathway that determines embryo pattern formation and morphogenesis. Prevents meiosis resumption in prophase-arrested oocytes via CDC25B inactivation by phosphorylation. May also regulate rapid eye movement (REM) sleep in the pedunculopontine tegmental (PPT). Phosphorylates APOBEC3G and AICDA. Isoform 2 phosphorylates and activates ABL1 in sperm flagellum to promote spermatozoa capacitation. -
Tissue specificity
Isoform 1 is ubiquitous. Isoform 2 is sperm-specific and is enriched in pachytene spermatocytes but is not detected in round spermatids. -
Sequence similarities
Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. cAMP subfamily.
Contains 1 AGC-kinase C-terminal domain.
Contains 1 protein kinase domain. -
Post-translational
modificationsAsn-3 is partially deaminated to Asp giving rise to 2 major isoelectric variants, called CB and CA respectively.
Autophosphorylated. Phosphorylation is enhanced by vitamin K(2). Phosphorylated on threonine and serine residues. Phosphorylation on Thr-198 is required for full activity.
Phosphorylated at Tyr-331 by activated receptor tyrosine kinases EGFR and PDGFR; this increases catalytic efficienncy. -
Cellular localization
Cytoplasm. Cell membrane. Nucleus. Mitochondrion. Translocates into the nucleus (monomeric catalytic subunit). The inactive holoenzyme is found in the cytoplasm. Distributed throughout the cytoplasm in meiotically incompetent oocytes. Associated to mitochondrion as meiotic competence is acquired. Aggregates around the germinal vesicles (GV) at the immature GV stage oocytes and Cell projection, cilium, flagellum. Expressed in the midpiece region of the sperm flagellum. - Information by UniProt
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Database links
- Entrez Gene: 5566 Human
- Entrez Gene: 18747 Mouse
- Entrez Gene: 25636 Rat
- Omim: 601639 Human
- SwissProt: P17612 Human
- SwissProt: P05132 Mouse
- SwissProt: P27791 Rat
- Unigene: 631630 Human
see all -
Alternative names
- cAMP dependent protein kinase alpha catalytic subunit antibody
- cAMP dependent protein kinase beta catalytic subunit antibody
- cAMP dependent protein kinase catalytic beta subunit isoform 4ab antibody
see all
Images
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All lanes : Anti-cAMP Protein Kinase Catalytic subunit antibody [EP2102Y] (ab76238) at 1/20000 dilution (Purified)
Lane 1 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
Lane 3 : C6 (Rat glial tumor glial cell) whole cell lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 46 kDa
Observed band size: 42 kDa why is the actual band size different from the predicted? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat stomach tissue sections labeling cAMP Protein Kinase Catalytic subunit with purified ab76238 at 1/750 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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ab76238 (purified) at 1/40 dilution (2 µg) immunoprecipitating cAMP Protein Kinase Catalytic subunit in MCF-7 whole cell lysate.
Lane 1 (input): MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10 µg
Lane 2 (+): ab76238 & MCF-7 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76238 in MCF-7 whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
Intracellular Flow Cytometry analysis of MCF-7 (Human breast adenocarcinoma epithelial cell) cells labeling cAMP Protein Kinase Catalytic subunit with purified ab76238 at 1/80 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling cAMP Protein Kinase Catalytic subunit with purified ab76238 at 1:100 dilution (8.0 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebrum tissue sections labeling cAMP Protein Kinase Catalytic subunit with purified ab76238 at 1/750 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid carcinoma tissue sections labeling cAMP Protein Kinase Catalytic subunit with purified ab76238 at 1/750 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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All lanes : Anti-cAMP Protein Kinase Catalytic subunit antibody [EP2102Y] (ab76238) at 1/200000 dilution (unpurified)
Lane 1 : HeLa cell lysate
Lane 2 : MCF-7 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : goat anti-rabbit HRP at 1/1000 dilution
Predicted band size: 46 kDa
Observed band size: 42 kDa why is the actual band size different from the predicted? -
ab76238 (unpurified), at a 1/100 dilution, staining human cAMP Protein Kinase Catalytic Subunit in testis by Immunohistochemistry, Formalin/PFA-fixed paraffin-embedded tissue. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
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ICC/IF image of ab76238 (unpurified) stained HeLa cells. The cells were 4% formaldehyde (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab76238, neat) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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Overlay histogram showing HeLa cells stained with ab76238 (unpurified) (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76238, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (26)
ab76238 has been referenced in 26 publications.
- Wen J et al. Role of transient receptor potential vanilloid-1 in behavioral thermoregulation of the Mongolian gerbil Meriones unguiculatus. Integr Zool 17:608-618 (2022). PubMed: 34498418
- Yu S et al. BMS-470539 Attenuates Oxidative Stress and Neuronal Apoptosis via MC1R/cAMP/PKA/Nurr1 Signaling Pathway in a Neonatal Hypoxic-Ischemic Rat Model. Oxid Med Cell Longev 2022:4054938 (2022). PubMed: 35140838
- Zhan Y et al. Effects of Maren Pills on the Intestinal Microflora and Short-Chain Fatty Acid Profile in Drug-Induced Slow Transit Constipation Model Rats. Front Pharmacol 13:804723 (2022). PubMed: 35496291
- Pagano Zottola AC et al. Expression of Functional Cannabinoid Type-1 (CB1) Receptor in Mitochondria of White Adipocytes. Cells 11:N/A (2022). PubMed: 36010658
- Wan MX et al. Integrating network pharmacology and experimental verification to explore the mechanism of puerarin against oliguria in acute alcoholism. Front Pharmacol 13:1006660 (2022). PubMed: 36299877