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 16 publications.
- Shahrestanaki MK et al. IPP-1 controls Akt/CREB phosphorylation extension in A2a adenosine receptor signaling cascade in MIN6 pancreatic ß-cell line. Eur J Pharmacol 850:88-96 (2019). PubMed: 30772395
- Chen WC et al. Xenotropic and polytropic retrovirus receptor 1 (XPR1) promotes progression of tongue squamous cell carcinoma (TSCC) via activation of NF-?B signaling. J Exp Clin Cancer Res 38:167 (2019). PubMed: 30995931
- Hurtado Del Pozo C et al. A Receptor of the Immunoglobulin Superfamily Regulates Adaptive Thermogenesis. Cell Rep 28:773-791.e7 (2019). PubMed: 31315054
- Mandelbaum AD et al. miR-17-92 and miR-106b-25 clusters regulate beta cell mitotic checkpoint and insulin secretion in mice. Diabetologia 62:1653-1666 (2019). PubMed: 31187215
- Chen R et al. cAMP metabolism controls caspase-11 inflammasome activation and pyroptosis in sepsis. Sci Adv 5:eaav5562 (2019). PubMed: 31131320