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ProteomicsPRM analysis of phosphopeptides of Nab2, Foxk1, HURP, ERK1/2, NPM, and Hspa4 from ΔB-Raf:ER cells.
ΔB-Raf:ER cells were treated with U0126 or 4-HT for 30 min in four biological replicates. Total proteins from cell lysates were digested with Typsin/Lys-C mix, and phosphopeptides were enriched by IMAC using Fe-NTA columns. After data-dependent LC-MS/MS analysis, phosphopeptides of Nab2, Foxk1, HURP...
ORGANISM(S): Mus Musculus (mouse) 
2022-03-03 | PXD028826 | JPOST Repository
ProteomicsPhos-tag SDS-PAGE followed by LC-MS/MS analysis of in vitro phosphorylated Nab2, Foxk1, and HURP
Purified GST-fused Nab2, Foxk1, and HURP were incubated with ATP in the presence or absence of His-tagged active ERK in vitro, separated by Phos-tag SDS-PAGE, and visualized by CBB staining. The most shifted regions of protein bands on the Phos-tag-containing gels were excised, digested with trypsin...
ORGANISM(S): Mus Musculus (mouse) 
2022-03-03 | PXD028837 | JPOST Repository
ProteomicsLabel-free quantitative phosphoproteomics of ΔB-Raf:ER cells treated with U0126 or 4-HT
ΔB-Raf:ER cells were treated with 10 µM U0126 or 1 µM 4-HT for 30 min. The cells were lysed in 6 M guanidine-HCl containing 100 mM HEPES-NaOH (pH7.5), 10 mM TCEP, and 40 mM CAA. The lysates were dissolved by heating and sonication and then centrifuged. The supernatants were recovered, and proteins (...
ORGANISM(S): Mus Musculus (mouse) 
2022-03-03 | PXD028723 | JPOST Repository
ProteomicsTMTpro-18plex analysis of MM1.S cells treated with thalidomide and its derivatives
MM1.S cells were treated with DMSO or 10 µM thalidomide or its derivatives for 5 h in two biological replicates. The cells were lysed in 200 µL guanidine buffer (6 M guanidine-HCl, 100 mM HEPES-NaOH, pH 7.5, 10 mM TCEP, 40 mM chloroacetamide). After heating and sonication, proteins (100 µg each) wer...
ORGANISM(S): Homo Sapiens (human) 
2023-07-22 | PXD041812 | JPOST Repository
ProteomicsSequential Digestion with Different Proteases
We examined three combinations of different proteases for digestion of proteins from TurboID-STING-expressing cells: trypsin alone, Lys-C followed by trypsin, and Glu-C followed by trypsin. First, we tested whether PTS buffer used to solubilize precipitated proteins is compatible with digestion with...
ORGANISM(S): Mus Musculus (mouse) 
2022-08-25 | PXD035242 | JPOST Repository
ProteomicsTMTpro-18plex analysis of siSTK38 or siLuc HeLa cells treated with EBSS or LLOMe
HeLa cells were transfected with STK38 siRNA or Luc siRNA and then treated with EBSS or LLOMe in three biological replicates. The cells were lysed in 500 µL guanidine buffer (6 M guanidine-HCl, 100 mM HEPES-NaOH, pH 7.5, 10 mM TCEP, 40 mM chloroacetamide). After heating and sonication, proteins (330...
ORGANISM(S): Homo Sapiens (human) 
2023-11-21 | PXD039312 | JPOST Repository
ProteomicsTMTpro-18plex analysis of MM1.S cells treated with PROTACs
MM1.S cells were treated with DMSO, 0.3 µM Po-P, 0.3 µM Le-P, 0.3 µM F-P, 0.3 µM Cl-P, or 0.3 µM F3C-P for 16 h in three biological replicates. The cells were lysed in 100 µL guanidine buffer (6 M guanidine-HCl, 100 mM HEPES-NaOH, pH 7.5, 10 mM TCEP, 40 mM chloroacetamide). After heating and sonicat...
ORGANISM(S): Homo Sapiens (human) 
2023-07-22 | PXD037178 | JPOST Repository
ProteomicsTMTpro-18plex analysis of NTERA-2 cells treated with PROTACs
NTERA-2 cells were treated with DMSO, 0.3 µM Po-P, 0.3 µM Le-P, 0.3 µM F-P, 0.3 µM Cl-P, or 0.3 µM F3C-P for 16 h in three biological replicates. The cells were lysed in 100 µL guanidine buffer (6 M guanidine-HCl, 100 mM HEPES-NaOH, pH 7.5, 10 mM TCEP, 40 mM chloroacetamide). After heating and sonic...
ORGANISM(S): Homo Sapiens (human) 
2023-07-22 | PXD037179 | JPOST Repository
ProteomicsOptimization of Elution Conditions for Biotinylated Peptides from Tamavidin 2-REV
We attempted to optimize the conditions for competitive elution of biotinylated peptides from the Tamavidin 2-REV beads. First, to optimize the temperature at which biotinylated peptides were eluted from the beads, biotinylated peptides were eluted with the biotin solution at 4, 37, 56, and 95 ºC. S...
ORGANISM(S): Mus Musculus (mouse) 
2022-08-25 | PXD035219 | JPOST Repository
ProteomicsPurification of Biotinylated Peptides using SDB-SCX StageTip
We tried purification of biotinylated peptides using SDB-SCX StageTip to reduce interference ions. We compared this method with the original and optimized workflows. We assessed reproducibility of each enrichment process by three technical replicates. Original workflow with SDB-SCX: A 15-µL slurry o...
ORGANISM(S): Mus Musculus (mouse) 
2022-08-25 | PXD035229 | JPOST Repository
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