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Defining the cellular response to perturbations is crucial for further illuminating the functionality of the phosphoproteome. Here we describe \u00b5Phos (\u2018microPhos\u2019), an accessible phosphoproteomics platform that permits phosphopeptide enrichment from 96-well cell culture and small tissue amounts in &lt;8\u2009h total processing time. By greatly minimizing transfer steps and liquid volumes, we demonstrate increased sensitivity, &gt;90% selectivity, and excellent quantitative reproducibility. Employing highly sensitive trapped ion mobility mass spectrometry, we quantify ~17,000 Class I phosphosites in a human cancer cell line using 20\u2009\u00b5g starting material, and confidently localize ~6200 phosphosites from 1\u2009\u00b5g. This depth covers key signaling pathways, rendering sample-limited applications and perturbation experiments with hundreds of samples viable. 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