Proteins and synapses
Detection, measurement and manipulation of proteins, receptors and synapses: Western blotting, receptor pharmacology and synaptic plasticity.
- Co-immunoprecipitation of synaptic proteinsCo-immunoprecipitation to test whether the scaffold protein PSD-95 and the NMDA receptor subunit GluN2B are associated in brain tissue.Guided experiment
- Measuring protein concentration by sandwich ELISAMeasurement of the concentration of BDNF in two groups of mice or people by sandwich ELISA, from sample preparation to the interpretation of the standard curve.Guided experiment
- Counting dendritic spines with Golgi–Cox stainingA test of whether dendritic spines are less dense on pyramidal neurons in one group than in another, by Golgi–Cox staining, blind spine counts and an analysis of nested data.Guided experiment
- Measuring how strongly a drug acts: concentration–responseThe potency and efficacy of a reference agonist and two coded test compounds are measured on cells that carry their receptor, from weighing the powder to a statistical comparison, and two antagonists are then classified by Schild analysis.Guided experiment
- Long-term potentiation in a hippocampal sliceSynaptic strength is recorded at the CA3 to CA1 synapses of a hippocampal slice, strengthened with a burst of high-frequency stimulation, and tested for its dependence on NMDA receptors, with the analysis a laboratory would use.Guided experiment
- Isolating synapses: synaptosome and postsynaptic density fractionationPurify synaptosomes and the postsynaptic density from mouse or human brain tissue, prove with marker proteins what each fraction contains, and ask whether a receptor subunit is redistributed to synapses in one group compared with another.Guided experiment
- Measuring a protein by Western blottingOne protein is measured in the hippocampus of two groups of mice, from frozen tissue to a statistical comparison: lysis, protein assay and loading, electrophoresis and transfer, antibodies and exposure, blind quantification and analysis.Guided experiment
- Western blotting: from brain tissue to quantified bandsLysis of brain tissue, a BCA protein assay, SDS-PAGE, transfer, probing and imaging of a simulated protein, and blind quantification of the bands across groups of animals.Simulator
- Receptor pharmacology in a cell-based assayConcentration–response curves of agonists and antagonists in a 96-well calcium-flux assay, with estimates of potency, efficacy and antagonist affinity.Simulator
- Synaptic plasticity in hippocampal slicesField EPSPs at CA3 to CA1 synapses, the induction of long-term potentiation or depression, and a test of its dependence on NMDA receptors.Simulator
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