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Picoprobe accoa assay kit

Manufactured by Abcam
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The PicoProbe AcCoA assay kit is a fluorometric assay designed to quantify Acetyl Coenzyme A (AcCoA) levels in biological samples. The kit utilizes a proprietary probe that specifically binds to AcCoA, generating a fluorescent signal proportional to the AcCoA concentration in the sample.

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Lab products found in correlation

Versamax tunable microplate reader, by Molecular Devices (2 mentions) Spark multimode microplate reader, by Tecan (1 mentions)

4 protocols using picoprobe accoa assay kit

1

Acetyl-CoA Quantification Assay

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AcCoA content was determined on total or cytosolic fractions using the PicoProbe AcCoA assay kit (ab87546, Abcam) according to manufacturer’s instructions. Briefly, after deproteinization using the perchloric acid, the CoASH Quencher and Quencher remover were added into the sample to correct the background generated by free CoASH and succ-CoA. The sample was then diluted with the reaction mix, and fluorescence was measured using a plate reader and the following settings: λex = 535 nm; λem = 587 nm. Fluorescence was measured using a Versamax Tunable microplate reader (Molecular Devices) or Spark multimode microplate reader (TECAN Trading AG, Switzerland). The acetyl-CoA standard curve was made in the range of 0–100 pM and the correlation coefficient was 0.990 or higher.
Son S.M., Park S.J., Stamatakou E., Vicinanza M., Menzies F.M, & Rubinsztein D.C. (2020). Leucine regulates autophagy via acetylation of the mTORC1 component raptor. Nature Communications, 11, 3148.
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2

Measuring Acetyl-CoA Levels

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The PicoProbe AcCoA assay kit (ab87546, Abcam, Cambridge, UK) was used to assess the AcCoA content in total or cytosolic fractions, following the manufacturer’s protocol.
Wei L., Wang R., Wazir J., Lin K., Song S., Li L., Pu W., Zhao C., Wang Y., Su Z, & Wang H. (2022). 2-Deoxy-D-glucose Alleviates Cancer Cachexia-Induced Muscle Wasting by Enhancing Ketone Metabolism and Inhibiting the Cori Cycle. Cells, 11(19), 2987.
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3

Ac-CoA Quantification Assay Protocol

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Ac‐CoA content was measured using the PicoProbe Ac‐CoA assay kit (Abcam, ab87546) according to manufacturer's instructions. Briefly, after deproteinization using the perchloric acid, the CoA‐SH Quencher and Quencher remover were added into the samples to correct the background generated by free CoA‐SH and succ‐CoA. The samples were then diluted with the reaction mixture, and the fluorescence of samples was measured using a multifunctional microplate reader (EnVision PE, USA) with the parameters settings of λex = 535 nm and λem = 587 nm. The Ac‐CoA standard curve was constructed in the range of 0–100 × 10−12m with the correlation coefficient of ≥0.99.
He Y., Wang S., Liu S., Qin D., Liu Z., Wang L., Chen X, & Zhang L. (2023). MSL1 Promotes Liver Regeneration by Driving Phase Separation of STAT3 and Histone H4 and Enhancing Their Acetylation. Advanced Science, 10(23), 2301094.
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4

Quantifying Bacterial Intracellular Acetyl-CoA

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Bacterial intracellular acetyl-coenzyme A (Ac-CoA) content was determined using the PicoProbe Ac-CoA assay kit (Abcam) according to manufacturer's instructions. Briefly, PA14 and the pvrA mutant were grown for 10 h in the Glu-M9 or FA-M9 medium. 30 ml of the bacterial cultures were collected by centrifugation at 12 000 × g for 5 min. After deproteinization with perchloric acid, the coenzyme A (CoA) Quencher and Quencher remover were added into each sample to correct the background generated by free CoA and succinyl-coenzyme A (succ-CoA). The sample was then diluted with the reaction mix, and the fluorescence was measured using a Versamax Tunable microplate reader (Molecular Devices) at the following settings: λex 535 nm; λem 587 nm. The acetyl-CoA standard curve was made in the range of 0–100 pM and the correlation coefficient was 0.990 or higher.
Pan X., Fan Z., Chen L., Liu C., Bai F., Wei Y., Tian Z., Dong Y., Shi J., Chen H., Jin Y., Cheng Z., Jin S., Lin J, & Wu W. (2020). PvrA is a novel regulator that contributes to Pseudomonas aeruginosa pathogenesis by controlling bacterial utilization of long chain fatty acids. Nucleic Acids Research, 48(11), 5967-5985.
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