Lactate colorimetric assay kit

To determine whether miR-130b is involved in glycolysis, cumulus cells were transfected with 50 nM of miR-130b precursor, inhibitor, or mismatch in 24-wells plate in serum-free medium. Twenty-four hours post-transfection, the concentration of lactate, the end product of glycolysis, was determined using the lactate colorimetric assay kit (Abcam, Cambridge, MA, USA). The OD was measured at 450 nm and the standard curve plot (nmol/well vs. OD 450 nm) was then generated. Finally, the lactate concentrations were determined as follows: C = La/Sv (nmol/μl or mM), where La is the lactic acid amount (nmol) and Sv is the sample volume (μl) in the well.

Lactate release was measured with the Lactate Colorimetric Assay Kit (cat# ab65331, Abcam, Cambridge, UK), according to manufacturer's indication. Briefly, a total of 10 × 10³ cells/well were plated in 24-well plate in the presence or absence of serum and processed as described previously 23 (link).

The concentration of lactate, the end product of glycolysis, was determined using a lactate colorimetric assay kit (Abcam, MA, USA). Cells were cultured in 6-well plates, and then harvested and deproteinized according to the manufacturer’s protocol. The optical density was measured at 450 nm and a standard curve plot (nmol/well vs. OD 450 nm) was generated using serial dilutions of lactate. Lactate concentrations were calculated with formula provided by the kit manufacturer.

Energy production, acidosis-related markers, and lipid peroxidation markers were validated to elucidate the neuroprotective effects of PEP-1-PGK1 against ischemic damage in the hippocampus of gerbils. Animals were sacrificed with isoflurane, and their hippocampi were dissected and homogenized in the assay buffer. Intracellular pH, ATP, lactate, and lipid peroxidation levels were measured using commercially available assay kits (a fluorometric intracellular pH assay kit [Merck, Darmstadt, Germany], ATP determination kit [Molecular Probes], lactate colorimetric assay kit [Abcam, Cambridge, UK], and MDA assay kit [Abcam], respectively), according to each manufacturer’s protocol.

A total of 2 × 10⁶ cells were treated for 24 h with 10 mM DCA and lactate was measured in culture medium using a Lactate Colorimetric Assay Kit (Abcam, Cambridge, MA, USA). Data were normalized to total protein amounts in each sample.

Lactate concentration was measured using the Lactate Colorimetric Assay Kit (Abcam, Cambridge, MA, USA). Equal number of cells were suspend in Lactate Assay Buffer and all experimental procedure was performed according to the instruction of the manufacturer. Experiment was repeated three times and absorbance was measured using the VERSA microplate reader (Turner Biosystems). For p53 inhibition, 20 μM pifithrin-α (PFT-α) (Sigma-Aldrich, MO, USA) was treated for 1 h before the cisplatin treatment and quantifications were made at indicated time points. For N-acetylcysteine (NAC) (Sigma-Aldrich), which is a powerful antioxidant, 10 μM NAC was pretreated for 1 h and then cisplatin was applied. All experiments related to PFT-α and NAC followed the same treatment process.