The timsTOF Pro 2 is a new proteomics workhorse for robust, unbiased, deep and quantitative 4D-proteomics and 4D-epiproteomics for plasma, tissue samples and from cell cultures. Further design advances combined with enhanced dda-PASEF, dia-PASEF and prm-PASEF methods deliver industry-leading performance, with unparalleled robustness and throughput. The timsTOF Pro 2 allows the detection of >6000 proteins and >60,000 peptides by dda-PASEF with 60 min gradients on 200 ng of digest. It also has new high sensitivity methods that enable very good proteome coverage with 10× less digest, down to 20 ng, for >3500 protein groups and >25,000 unique peptides using a 30 min gradient. dia-PASEF workflows on timsTOF Pro 2 include an improved interface for designing experiments, and can now identify ~8000 protein groups and 70,000 peptides in 60 min on 200 ng of digests.