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Estimation of ATP of the bacterial population by a luciferase assay.

AA Ajit Kumar Akela
AK Ashwani Kumar
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Mtb cells (OD600 = 1.0) were incubated with BDQ (0.35 μM) and DCCD (100 μM) along with the control (DMSO) for 3 h, and equal volumes of cells from all samples in triplicates were then taken and centrifuged at 6,000 rpm (6,641 rcf) for 5 min, followed by washing of cells with Tris-EDTA (TE) buffer. Five hundred microliters of TE buffer was added to the bacterial pellets. Cells were then lysed with a bead beater (8 cycles, with each consisting of a 30-s pulse at a speed of 6.5 m/s2 followed by a 5-min incubation at ice). The cell lysate was centrifuged at 10,000 rpm (9,384 rcf) for 10 min. ATP was estimated using an ATP bioluminescence assay kit (catalogue no. 11699695001; CLS II) according to the manufacturer’s instructions. Briefly, 50 μl of the supernatant was mixed with reaction buffer containing luciferase in 96-well plates, and bioluminescence was measured. Data were plotted using GraphPad Prism software.

Copyright and License information:  ©2021 Akela and Kumar.
This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.

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