The Phospho Explorer antibody array assay was performed by Full Moon Biosystems on patient-derived pediatric glioblastoma (SU-pcGBM2) cells. Cell lysates were prepared using Protein Extraction Kit (Full Moon BioSystems). Clear supernatant of the lysates was separated, biotinylated, and incubated with Phospho Explorer Antibody Arrays (Full Moon BioSystems) for two hours at room temperature. The array slides were washed with Wash Buffer (Full Moon BioSystems) and rinsed with DI water. The slides were then incubated with Cy3-Streptavidin for 45 minutes at room temperature, then washed, rinsed and dried. Arrays were scanned on GenePix Array Scanner (Molecular Devices). Image quantification was performed on GenePix Pro (Molecular Devices). Signal intensity data for each spot on the array was extracted from array images. Since there are two replicates printed for each antibody, the mean signal intensity of the replicates is determined. The data is then normalized to the median value (signal intensity) of all antibodies on the slide. Finally, fold change between Control Sample and Treatment Sample is determined using the normalized data (Treatment Sample’s signal is divided by Control Sample’s signal).
Phospho explorer antibody array assay
Manufactured by Full Moon BioSystems
The Phospho Explorer antibody array assay is a multiplex platform that allows for the simultaneous detection and quantification of the phosphorylation status of multiple protein targets in a single experiment. The assay utilizes an array of highly specific antibodies targeting known phosphorylation sites, enabling researchers to obtain a comprehensive profile of cellular signaling pathways.
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2 protocols using phospho explorer antibody array assay
1
Phospho Explorer Antibody Array for Pediatric Glioblastoma
2
Phospho Explorer Antibody Array for Pediatric Glioblastoma
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The Phospho Explorer antibody array assay was performed by Full Moon Biosystems on patient-derived pediatric glioblastoma (SU-pcGBM2) cells. Cell lysates were prepared using Protein Extraction Kit (Full Moon BioSystems). Clear supernatant of the lysates was separated, biotinylated, and incubated with Phospho Explorer Antibody Arrays (Full Moon BioSystems) for two hours at room temperature. The array slides were washed with Wash Buffer (Full Moon BioSystems) and rinsed with DI water. The slides were then incubated with Cy3-Streptavidin for 45 minutes at room temperature, then washed, rinsed and dried. Arrays were scanned on GenePix Array Scanner (Molecular Devices). Image quantification was performed on GenePix Pro (Molecular Devices). Signal intensity data for each spot on the array was extracted from array images. Since there are two replicates printed for each antibody, the mean signal intensity of the replicates is determined. The data is then normalized to the median value (signal intensity) of all antibodies on the slide. Finally, fold change between Control Sample and Treatment Sample is determined using the normalized data (Treatment Sample’s signal is divided by Control Sample’s signal).
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