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Detection of Signaling Effector-Complexes Downstream of BMP4 Using in situ PLA, a Proximity Ligation Assay
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Detection of Signaling Effector-Complexes Downstream of BMP4 Using in situ PLA, a Proximity Ligation Assay

Article DOI: 10.3791/2631-v • 12:53 min • March 3rd, 2011
March 3rd, 2011 •
1, 1
1Medical Research Council, Clinical Sciences Centre, Imperial College, Hammersmith Hospital

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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.

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Detection Signaling Effector-complexes BMP4 In Situ PLA Proximity Ligation Assay BMP Receptors Smad1/5/8 Effectors Smad4 Nucleus Transcription Factors Traditional Immuno-fluorescence Techniques Phospho-Smad Peptides Sensitivity High Background Gross Quantification Antibody Signal Intensity Photosensitive Fluorescent Protein Interactions Specificity Amplification Of DNA Surrogate Localized Signal Spots Recognition Event Measurement
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