Analyses of culture models
Culture models are analysed for different parameters to characterize and quantify the effect of a test substance.
1) Cell survival
- MTT
- Calcein-AM
- LDH
2) Necrosis
- Propidium Iodide
3) Apoptosis
- YOPRO, an apoptosis specific DNA intercalator
- Caspase-3 activity
- Mitochondrial membrane potential
- DNA condensation
4) ELISA
- Aß1-38, Aß1-40, Aß1-42
- sAPP-alpha, sAPP-beta
- Signal transduction molecules
- Inflammation markers (e.g. Prostaglandin E2)
5) Immunocytochemistry
- Neuronal markers (ß-III tubulin, neurofilament, MAP2, Tyrosine hydroxylase)
- Glial markers (GFAP, CD11b)
- Disease related markers (Tau, phosphorylated Tau, signalling molecules)
- Synaptic proteins (synaptophysin)
6) Western Blotting
- Synaptic proteins (Glutamate receptor 1, NMDA receptors, synaptophysin, drebrin) e.g. from matured hippocampal neurons
- Tau and phospho-Tau isoforms
- Aß1-42, Aß1-40 & other species, Aß aggregation assays
- Cell signalling molecules
7) Quantification of neurite outgrowth
- Area measurement (semi-automatic) using Image Pro Plus software (see Histology & Imaging)
- Neurite length using Image Pro Plus