Quantitative Determination of Lactate in Microglia Cells

Lactate content in the supernatant was measured using the EnzyChrom™ Glycolysis Assay Kit according to manufacturer’s protocol. Microglia were seeded in a 12-well plate at a density of 5x10⁴ cells per well. Cells were allowed to adhere overnight, then incubated in serum-free medium and treated with LPA 1 µM for indicated time periods. At the end of the treatments, the supernatants were collected, mixed with the provided enzyme mix and optical density was measured at 565 nm to quantify lactate content.

These analyses revealed that the media concentrations of lactate were significantly higher in BV-2 cells exposed to LPA for 8 and 24 h, indicating a shift toward aerobic glycolysis.

The kit works as expected and gives reproducible results. Based on our experience, it is better (when possible) to use the supernatants immediately and not freezing and thawing them in order to perform the analysis at a later stage.