(b) Aperture 384 Plate US Luminescence aperture 9, height
4.2 mm, diameter 3.7 mm.
25. We recommended generating standard curves for OD vs CFUs
and Luminescence vs CFU during the setup of the protocol
with your conditions and luminometer and spectrophotometer. It is possible to use pre-counted glycerol aliquots stored at
À80
C up to 3 months instead of fresh mycobacterial culture
for infecting the monocytes.
References
1. Global tuberculosis report 2019 (2019) World
Health Organization, Geneva. Licence: CC
BY-NC-SA 3.0 IGO
2. Ballel L, Bates RH, Young RJ, Alvarez-GomezD, Alvarez-Ruiz E, Barroso V, Blanco D,
Crespo B, Escribano J, Gonzalez R,
Lozano S, Huss S, Santos-Villarejo A, MartinPlaza JJ, Mendoza A, Rebollo-Lopez MJ,
Remuin ˜an-Blanco M, Lavandera JL, PerezHerran E, Gamo-Benito FJ, Garcia-Bustos JF,
Barros D, Castro JP, Cammack N (2013) Fueling open-source drug discovery: 177 smallmolecule leads against tuberculosis. Chem
Med Chem 8:313–321
3. Sorrentino F, Gonzalez-Del-Rio R, Zheng X,
Presa-Matilla J, Torres-Gomez P, MartinezHoyos M, Pe ´rez-Herra ´n ME, MendozaLosana A, Av-Gay Y (2016) Development of
an intracellular screen for new compounds able
to inhibit Mycobacterium tuberculosis growth in
human macrophages. Antimicrob Agents Chemother 60:640–645
4. Home DJ, Pinto LM, Arentz M, Lin SY,
Desmond E, Flores LL, Steingart KR, Minion
J (2013) Diagnostic accuracy and reproducibility of WHO-endorsed phenotypic drug susceptibility testing methods for first-line and
second-line antituberculosis drugs. J Clin
Microbiol 51:393–401
5. Franzblau SG, Witzig RS, McLaughlin JC,
Torres P, Madico G, Hernandez A, Degnan
MT, Cook MB, Quenzer VK, Ferguson RM,
Gilman RH (1998) Rapid, low-technology
MIC determination with clinical Mycobacterium tuberculosis isolates by using the microplate Alamar Blue assay. J Clin Microbiol
36:362–366
6. Fan F, Buttler B, Riss T, Wook K (2004) Quantitate microbial cells using a rapid and sensitive
ATP-bases luminescent assay. Promega Notes
88:2–4
7. Fan F, Buttler B, Riss T, Wook K (2005)
BacTiter-Glo assay for antimicrobial drug discovery and general microbiology. Promega
Notes 89:25–27
8. Wodnicka M, Guarino RD, Hemperly JJ, Timmins MR, Stitt D, Pitner JB (2000) Novel
fluorescent technology platform for high
throughput cytotoxicity and proliferation
assays. J Biomol Screen 5:141–152
9. Rivera-Marrero CA, Stewart J, Shafer WM,
Roman J (2004) The down-regulation of
cathepsin G in THP-1 monocytes after infection with Mycobacterium tuberculosis is associated with increased intracellular survival of
bacilli. Infect Immun 72:5712–5721
10. Bright-Glo™ luciferase assay system. Technical
bulletin. Promega
648
Esther Pe ´ rez-Herra ´ n and Alfonso Mendoza
4.2 mm, diameter 3.7 mm.
25. We recommended generating standard curves for OD vs CFUs
and Luminescence vs CFU during the setup of the protocol
with your conditions and luminometer and spectrophotometer. It is possible to use pre-counted glycerol aliquots stored at
À80
C up to 3 months instead of fresh mycobacterial culture
for infecting the monocytes.
References
1. Global tuberculosis report 2019 (2019) World
Health Organization, Geneva. Licence: CC
BY-NC-SA 3.0 IGO
2. Ballel L, Bates RH, Young RJ, Alvarez-GomezD, Alvarez-Ruiz E, Barroso V, Blanco D,
Crespo B, Escribano J, Gonzalez R,
Lozano S, Huss S, Santos-Villarejo A, MartinPlaza JJ, Mendoza A, Rebollo-Lopez MJ,
Remuin ˜an-Blanco M, Lavandera JL, PerezHerran E, Gamo-Benito FJ, Garcia-Bustos JF,
Barros D, Castro JP, Cammack N (2013) Fueling open-source drug discovery: 177 smallmolecule leads against tuberculosis. Chem
Med Chem 8:313–321
3. Sorrentino F, Gonzalez-Del-Rio R, Zheng X,
Presa-Matilla J, Torres-Gomez P, MartinezHoyos M, Pe ´rez-Herra ´n ME, MendozaLosana A, Av-Gay Y (2016) Development of
an intracellular screen for new compounds able
to inhibit Mycobacterium tuberculosis growth in
human macrophages. Antimicrob Agents Chemother 60:640–645
4. Home DJ, Pinto LM, Arentz M, Lin SY,
Desmond E, Flores LL, Steingart KR, Minion
J (2013) Diagnostic accuracy and reproducibility of WHO-endorsed phenotypic drug susceptibility testing methods for first-line and
second-line antituberculosis drugs. J Clin
Microbiol 51:393–401
5. Franzblau SG, Witzig RS, McLaughlin JC,
Torres P, Madico G, Hernandez A, Degnan
MT, Cook MB, Quenzer VK, Ferguson RM,
Gilman RH (1998) Rapid, low-technology
MIC determination with clinical Mycobacterium tuberculosis isolates by using the microplate Alamar Blue assay. J Clin Microbiol
36:362–366
6. Fan F, Buttler B, Riss T, Wook K (2004) Quantitate microbial cells using a rapid and sensitive
ATP-bases luminescent assay. Promega Notes
88:2–4
7. Fan F, Buttler B, Riss T, Wook K (2005)
BacTiter-Glo assay for antimicrobial drug discovery and general microbiology. Promega
Notes 89:25–27
8. Wodnicka M, Guarino RD, Hemperly JJ, Timmins MR, Stitt D, Pitner JB (2000) Novel
fluorescent technology platform for high
throughput cytotoxicity and proliferation
assays. J Biomol Screen 5:141–152
9. Rivera-Marrero CA, Stewart J, Shafer WM,
Roman J (2004) The down-regulation of
cathepsin G in THP-1 monocytes after infection with Mycobacterium tuberculosis is associated with increased intracellular survival of
bacilli. Infect Immun 72:5712–5721
10. Bright-Glo™ luciferase assay system. Technical
bulletin. Promega
648
Esther Pe ´ rez-Herra ´ n and Alfonso Mendoza