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Signatect pka assay system

Manufactured by Promega
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The SignaTECT PKA assay system is a laboratory equipment used for the detection and quantification of protein kinase A (PKA) activity. It provides a standardized and reproducible method for measuring PKA activity in biological samples.

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Lab products found in correlation

Complete dmem media, by Thermo Fisher Scientific (1 mentions)

2 protocols using signatect pka assay system

1

Measuring PKA Activity in H9c2 Cells

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H9c2 primary heart cells (ATCC, Manassas, VA) were plated in 6-well plates at 2.5 million cells/well overnight in complete DMEM media (Thermo Fisher, Waltham, MA, USA), at 37 °C with 5% CO2. An assay was performed as previously described [50 (link)]. In short, the following day, rabbit serum (1/100 dilution) from placebo and immunized groups were incubated separately with heart cell line in a final volume of 2 mL of medium. The cells were mechanically dislodged, centrifuged, and solubilized in extraction buffer. Protein kinase A (PKA) activity was measured with a SignaTECT PKA assay system (Promega, Madison, WI) per manufacturer’s instructions. The PKA activity is reported as a percent above the basal level.
Troese M.J., Burlet E., Cunningham M.W., Alvarez K., Bentley R., Thomas N., Carwell S, & Morefield G.L. (2023). Group A Streptococcus Vaccine Targeting the Erythrogenic Toxins SpeA and SpeB Is Safe and Immunogenic in Rabbits and Does Not Induce Antibodies Associated with Autoimmunity. Vaccines, 11(9), 1504.
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2

Determination of PKA Activity in HT22 Cells

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The PKA activity was determined using the SignaTECT PKA assay system (Promega), as described previously [17 (link), 36 (link), 37 (link)]. HT22 cells were pretreated without or with H89 and then treated in the absence or presence of different retinoids. Cells were then washed with 0.01M cold PBS and collected in an extraction buffer with protease inhibitor cocktails. Cells were then sonicated, centrifuged at 14,000 × g for 5 min at 4°C, and the supernatant (5μl from each group) was added to 20μl of kinase reaction mixture and incubated for 5 min at RT. The reaction was stopped by adding 12μl of termination buffer and samples were processed for determining the PKA activity [17 (link), 36 (link)].
Manna P.R., Reddy A.P., Pradeepkiran J.A., Kshirsagar S, & Reddy P.H. (2022). Regulation of retinoid mediated StAR transcription and steroidogenesis in hippocampal neuronal cells: Implications for StAR in protecting Alzheimer’s disease. Biochimica et biophysica acta. Molecular basis of disease, 1869(2), 166596.
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