Key features and details
- Assay type: Enzyme activity
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 4 hr 30 min
- Sample type: Adherent cells, Purified protein, Suspension cells, Tissue Extracts
Product namePKA Kinase Activity Assay Kit
Intra-assay Sample n Mean SD CV% Sample < 10% Inter-assay Sample n Mean SD CV% Sample <=12%
Sample typeAdherent cells, Suspension cells, Tissue Extracts, Purified protein
Assay typeEnzyme activity
Assay time4h 30m
PKA Kinase Activity Kit (ab139435) is a non-radioactive assay providing a safe, rapid and reliable method for the screening of inhibitors or activators of PKA and for quantitating the activity of PKA in purified or partially purified enzyme preparations. This kit is based on a solid phase enzyme-linked immuno-absorbent assay (ELISA) that utilizes a specific synthetic peptide as a substrate for PKA and a polyclonal antibody that recognizes the phosphorylated form of the substrate. The assay is designed for the analysis of PKA activity in the solution phase. This kit gives you the ability to an end-point or kinetic assay read-out in a convenient 96-well plate based assay and offers easy sample handling protocols.
The kit offers the following advantages:
1. Safe - non-radioactive measurement of kinase activity
2. Flexible - kinetic and end-point options available
3. Fast - results in < 4.5 hours
4. Efficient - only 30 µl diluted sample needed per well
Storage instructionsPlease refer to protocols.
Components 1 x 96 tests 20X Wash Buffer 1 x 30ml Active PKA 20 ng/µl 1 x 30µl Antibody Dilution Buffer 1 x 10ml Anti-Rabbit IgG: HRP Conjugate 1 x 20µl ATP 1 x 2mg Kinase Assay Dilution Buffer 1 x 10ml PKA Phosphospecific Substrate Antibody 1 x 5ml PKA Substrate Microtiter Plate 1 unit Stop Solution 2 1 x 10ml TMB Substrate 1 x 10ml
RelevanceIn cell biology, Protein kinase A (PKA) is a family of enzymes whose activity is dependent on cellular levels of cyclic AMP (cAMP). PKA is also known as cAMP-dependent protein kinase. Protein kinase A has several functions in the cell, including regulation of glycogen, sugar, and lipid metabolism.
1e7 3T3-L1 cells were treated for 15 minutes with 1 μM dexamethasone (ab120743) and 1 μg/mL insulin (ab123768) with the addition of 50 μM forskolin (ab120058), 0.5 mM IBMX (ab120840) or 0.1 mM 8-CPT-cAMP (ab120424). Cell extracts were prepared according to the protocol. Lysates were diluted 50-fold, and tested in duplicates (+/-SD).
Titration of positive control (active PKA), background signal subtracted (duplicates; +/- SD).
Titration of cAMP Protein Kinase (ab56268), background signal subtracted (duplicates; +/- SD).
ab139435 has been referenced in 25 publications.
- Zhao J et al. Upregulation of histamine receptor H1 promotes tumor progression and contributes to poor prognosis in hepatocellular carcinoma. Oncogene 39:1724-1738 (2020). PubMed: 31740780
- Wen Z et al. APPL1 knockdown blocks adipogenic differentiation and promotes adipocyte lipolysis. Mol Cell Endocrinol 506:110755 (2020). PubMed: 32045627
- Kitchen P et al. Targeting Aquaporin-4 Subcellular Localization to Treat Central Nervous System Edema. Cell 181:784-799.e19 (2020). PubMed: 32413299
- Lim J et al. Glutathione dynamics determine the therapeutic efficacy of mesenchymal stem cells for graft-versus-host disease via CREB1-NRF2 pathway. Sci Adv 6:eaba1334 (2020). PubMed: 32490200
- Chu S et al. Sinigrin Enhanced Antiasthmatic Effects of Beta Adrenergic Receptors Agonists by Regulating cAMP-Mediated Pathways. Front Pharmacol 11:723 (2020). PubMed: 32508648