Dataset Information

Towards sensitive, streamlined high throughput protein thermal stability analysis for low sample input

ABSTRACT: This method development aims to enhance and streamline excisting PISA workflows by reducing pipetting steps and digestion method. With this workflow we want to provide a method with higher reproducibility and simplicity. This method combines the excisting OneTip protocol with low cell concentration input on a 96 well format to reach a high kinase inhibitor target screening throughput from treatment to sample injection within 24h.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hela Cell

SUBMITTER: Maico Yannic Lechner

LAB HEAD: Jesper Velgaard Olsen

PROVIDER: PXD061105 | Pride | 2025-10-06

REPOSITORIES: Pride

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			Action	DRS
	20250129_AST5_Evo5_IO5cm_ML_80SPD_2Th2-5ms_Prot_HeLa_OTPISA_AKT_1uM_01.raw	Raw
	20250129_AST5_Evo5_IO5cm_ML_80SPD_2Th2-5ms_Prot_HeLa_OTPISA_AKT_1uM_02.raw	Raw
	20250129_AST5_Evo5_IO5cm_ML_80SPD_2Th2-5ms_Prot_HeLa_OTPISA_AKT_1uM_03.raw	Raw
	20250129_AST5_Evo5_IO5cm_ML_80SPD_2Th2-5ms_Prot_HeLa_OTPISA_AKT_1uM_04.raw	Raw
	20250129_AST5_Evo5_IO5cm_ML_80SPD_2Th2-5ms_Prot_HeLa_OTPISA_AKT_5uM_01.raw	Raw

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Publications

High-throughput proteome integral solubility alteration assay for low cell input using One-Tip.

Lechner Maico M Sabatier Pierre P Olsen Jesper V JV

Communications chemistry 20250926 1

Mass spectrometry-based versions of the cellular thermal shift assay (CETSA), like proteome integral solubility alteration (PISA), enable simultaneous monitoring of thousands of proteins for drug-target engagement. These methods are constrained in throughput and scalability, while the sample requirement limits the applicability to widely available material. Here, we combine PISA with the One-Tip method to simplify and streamline sample preparation. Using the mass spectrometry-compatible n-Dodecy ...[more]

PMID: 41006785

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