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Chapters
- 00:00Title
- 02:56Plating of Cells and Treatment with BMP
- 04:12Fixation and Permeabilization
- 05:26Blocking and Primary Antibodies Application
- 06:49The Proximity Ligation Assay (PLA)
- 09:54Preparation for Imaging
- 10:56In Situ Proximity Ligation Assay Results from Neuro2a Cells Treated with BMP4
- 12:18Conclusion
Here we show how to use Proximity Ligation Assay (PLA), with a combination of antibodies to visualize Bone Morphogenetic Protein (BMP) signaling in fixed cells. This technique allowed us to follow the nuclear accumulation of endogenous BMP activated effector-complexes and quantify their levels over time under BMP4 stimulation.
Tags
DetectionSignaling Effector-complexesBMP4In Situ PLAProximity Ligation AssayBMP ReceptorsSmad1/5/8 EffectorsSmad4NucleusTranscription FactorsTraditional Immuno-fluorescence TechniquesPhospho-Smad PeptidesSensitivityHigh BackgroundGross QuantificationAntibody SignalIntensityPhotosensitive FluorescentProtein InteractionsSpecificityAmplification Of DNA SurrogateLocalized SignalSpotsRecognition EventMeasurement
