Biochemistry88rigor
Single-Cycle Kinetics SPR Measurement of Antibody-Antigen Binding Affinity on a Biacore CM5 Sensor
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.
Immunology87rigor
Multiplex ELISA Optimization for Simultaneous Quantification of IL-6, TNF-α, and IFN-γ Secretion from LPS- and IFN-γ-Stimulated Human Macrophages
Optimize a multiplex ELISA cytokine panel to quantify IL-6, TNF-α, and IFN-γ secretion from stimulated macrophages.
Pharmacology84rigor
Cellular Thermal Shift Assay (CETSA) for Intracellular Target Engagement of Small-Molecule Inhibitors in Intact A549 Cells
Confirm direct, intracellular target engagement of a small-molecule inhibitor by measuring ligand-induced thermal stabilization of the endogenous target protein in intact A549 cells, generating melt curves and an isothermal dose-response (ITDRF) to estimate cellular engagement potency.
Structural Biology91rigor
Differential Scanning Fluorimetry (Thermofluor) Buffer and Ligand Stability Screen for a Purified Protein to Guide Structural Sample Preparation
Rank buffer, salt, pH, additive, and ligand conditions by their effect on protein thermal stability using SYPRO Orange differential scanning fluorimetry, identifying conditions that maximize stability for crystallization and cryo-EM sample preparation.