A high-capacity screen for adipogenic differentiation.
Glycerol-3-phosphate dehydrogenase (GPDH) is highly expressed in mature adipocytes. Activity of this enzyme is therefore routinely measured to assess adipogenic differentiation in cell cultures. Existing protocols for GPDH assays require relatively large amounts of cells, and throughput is limited due to multiple steps needed for cell harvest and enzyme extraction. We present here a new protocol allowing GPDH determinations to be performed in a 96-well-plate format. From the start of cell culture to the final readout all steps are carried out using the same multiwell plate, with a minimum of handling required. Our method is suitable for setting up high-throughput assays of adipogenic differentiation.
Sottile
V
V
Department of Research, Bone Metabolism, Novartis Pharma AG, CH-4002 Basel, Switzerland.
Seuwen
K
K
D017667:Adipocytes / Q000187:drug effects* / Q000201:enzymology* / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured / Q000187:drug effects / Q000502:physiology*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D007004:Hypoglycemic Agents / Q000494:pharmacology*; D009243:NAD / Q000737:chemistry / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear / Q000819:agonists; D000077154:Rosiglitazone; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*; D013844:Thiazoles / Q000494:pharmacology*; D045162:Thiazolidinediones; D014157:Transcription Factors / Q000819:agonists; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D053782:Transforming Growth Factor beta3
D007004:Hypoglycemic Agents; D018160:Receptors, Cytoplasmic and Nuclear; D013844:Thiazoles; D045162:Thiazolidinediones; D014157:Transcription Factors; D016212:Transforming Growth Factor beta; D053782:Transforming Growth Factor beta3; D000077154:Rosiglitazone; D009243:NAD; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases