Determine the association rate (ka), dissociation rate (kd), and equilibrium dissociation constant (KD) of a monoclonal antibody Fab fragment binding its protein antigen using single-cycle kinetics surface plasmon resonance, with the antibody captured via an anti-Fc surface and antigen injected as the analyte.
Determine the steady-state kinetic parameters (Km, Vmax, kcat, kcat/Km) of purified alkaline phosphatase by continuously monitoring p-nitrophenol release at 405 nm across a defined substrate concentration series, and quantify the inhibition mode and Ki of inorganic phosphate.
Purify a soluble His6-tagged recombinant protein to >90% homogeneity from E. coli BL21(DE3) using nickel-affinity chromatography, recovering sufficient yield (target >2 mg per liter culture) for downstream biophysical and enzymatic characterization.
Quantify mitochondrial respiratory parameters (basal respiration, ATP-linked respiration, maximal respiration, spare respiratory capacity, proton leak) in C2C12 myoblasts to detect oxidative phosphorylation defects under a metabolic perturbation.
Quantify glucose uptake capacity of resting versus TCR-activated primary human CD4+ T cells using the fluorescent glucose analog 2-NBDG by flow cytometry, to link T-cell activation state to glycolytic demand.
Use SYPRO Orange differential scanning fluorimetry (DSF/thermal shift) to determine the melting temperature (Tm) of a purified protein kinase domain and identify buffer conditions and small-molecule ligands that thermally stabilize it (positive ΔTm), guiding formulation and confirming ligand engagement.
Determine the absolute molar mass, oligomeric state, monodispersity, and aggregate content of a purified protein in solution using analytical size-exclusion chromatography coupled to in-line multi-angle light scattering and refractive-index detection (SEC-MALS), independent of column calibration standards.
Quantify glycolytic and TCA-cycle metabolite pools (glucose-6-phosphate, lactate, pyruvate, citrate, succinate, malate, α-ketoglutarate, etc.) in HepG2 cells by targeted hydrophilic interaction LC-MS/MS to characterize central carbon flux remodeling under treatment.
Quantify steady-state intracellular ATP (luciferase luminescence) and secreted/intracellular L-lactate (coupled enzymatic assay) in HeLa cells under normoxia versus hypoxia to assess the bioenergetic shift toward glycolytic ATP production.
Measure the extracellular acidification rate (ECAR) of MDA-MB-231 cells through the Glycolysis Stress Test (glucose, oligomycin, 2-deoxyglucose) to quantify basal glycolysis, glycolytic capacity, and glycolytic reserve in a highly glycolytic tumor line.