The BacTiter-Glo™ Microbial Cell Viability Assay provides a method for determining the number of viable microbial cells in culture based on quantitation of the ATP present. The homogeneous assay involves adding a single reagent (BacTiter-Glo™ Reagent) directly to bacterial cells cultured in medium and measuring luminescence. This assay format reduces pipetting errors that may be introduced during the multiple steps required of other ATP measurement methods.
The formulation of the reagent supports bacterial cell lysis and generation of a luminescent signal. The luminescent signal is proportional to the amount of ATP present, which is directly proportional to the number of viable cells in culture. The assay relies on the properties of a proprietary thermostable luciferase (Ultra-Glo™ Recombinant Luciferase) and a proprietary buffer formulation for extracting ATP from bacteria. The assay has been shown to detect a variety of bacteria and fungi.
Data can be recorded in 5 minutes or less after adding reagent and mixing. The assay's superior sensitivity allows the detection of growth or toxicity quickly after inoculation.
ATP can be measured from as few as 10 bacterial cells, and data is recorded by luminometer or CCD camera. The assay is 1,000-fold more sensitive than assays requiring absorbance readings. The "glow-type" luminescent signal is stable with a half-life generally greater than 30 minutes.
Applications of the assay include examining antimicrobial compound activity, screening antimicrobial compounds, examining bacterial growth with extended sensitivity and range, rapid bacterial quantitation, and rapid ATP detection.
Notes:
Using 100μl of BacTiter-Glo™ Reagent per assay in a 96-well format, Cat.# G8230 provides sufficient reagents to perform 100 assays; Cat.# G8231 and G8232, 1,000 assays; Cat.# G8233, 10,000 assays. Using 25μl of BacTiter-Glo™ Reagent per assay in a 384-well format, Cat.# G8230 provides sufficient reagents to perform 400 assays; Cat.# G8231 and G8232, 4,000 assays; Cat.# G8233, 40,000 assays. rATP (Cat.# P1132) must be purchased separately for performing a standard curve.