To analyze the signaling downstream of the b2AR in absence of different GRKs the eight different HEK293 cell lines with the genotypes CRISPR-Control, ΔQ-GRK, Δ2/3-GRK, Δ5/6-GRK, Δ3/5/6-GRK, Δ2/5/6-GRK, Δ2/3/6-GRK, Δ2/3/5-GRK were compared. The CRISPR-Control cell line possesses the endogenous activity of all GRKs, while the ΔQ-GRK has KOs for GRK2/3/5/6. The four cell lines Δ3/5/6-GRK, Δ2/5/6-GRK, Δ2/3/6-GRK and Δ2/3/5-GRK are deficient for three out of the fours GRKs of interest each and the two cell lines Δ2/3-GRK and Δ5/6-GRK are double KOs. The experiment was conducted in a 96-well plate format. The eight different HEK293 cell lines were either untreated control or treated with 10 µM isoprenaline for 5 min as concluded from the time resolved experiment. For each cell line and condition 6 replicates were made. Replicates were located next to each other on the plate due to sample handling requirements and later, before injection into the LC-MS, batch randomized in 8 batches with 16 samples each being unique in their combination of cell line and condition.