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ProteomicsTMT-based quantitative proteomics in Rab35 cKO P0 hippocampus
The hippocampi from the control (Rab35flox/+; Nestin-Cre) and Rab35 cKO mice groups at P0 (n = 5 biological replicates per genotype) were lysed in a buffer containing 6 M guanidine hydrochloride, 100 mM HEPES-NaOH (pH 8.0), 10 mM TCEP, and 40 mM chloroacetamide. The lysates were dissolved through he...
ORGANISM(S): Mus Musculus (mouse) 
2023-04-07 | PXD041290 | JPOST Repository
ProteomicsBioID2_BirA-MAVS_CLPB-BirA_PHB1-BirA_HEK293
HEK293 cells that stably express MAVS, CLPB or PHB1 fused at C-terminus with the biotin ligase from A. aeolicus were incubated with 100 μM biotin for 18 h. The mitochondrial fractions were lysed in RIPA buffer, and the clarified supernatants were incubated with streptavidin agarose beads for 2 h at ...
ORGANISM(S): Homo Sapiens (human) 
2019-09-05 | PXD014218 | JPOST Repository
ProteomicsBioID2_PHB2(V1)_PHB2(V3)-BirA-HA_HEK293
HEK293 cells that stably express PHB2 (V1 or V3) fused at C-terminus with the biotin ligase from A. aeolicus were incubated with 100 μM biotin for 18 h. The mitochondrial fractions were lysed in RIPA buffer, and the clarified supernatants were incubated with streptavidin agarose beads for 2 h at 4° ...
ORGANISM(S): Homo Sapiens (human) 
2019-09-05 | PXD011939 | JPOST Repository
ProteomicsBioID2_PHB2(V1)-(WT_KKK_I225P)
HEK293 cells that stably express wild-type (WT), E229/231/233K (KKK), or I225P mutant PHB2(V1) fused at C-terminus with the biotin ligase from A. aeolicus were incubated with 100 μM biotin for 18 h. The mitochondrial fractions were lysed in RIPA buffer, and the clarified supernatants were incubated ...
ORGANISM(S): Homo Sapiens (human) 
2019-09-05 | PXD011946 | JPOST Repository
ProteomicsComparison of Biotinylated Peptides Enriched by the Original and Optimized Workflows using Tamavidin 2-REV.
To evaluate the optimized workflow, biotinylated peptides enriched by the original and optimized workflows from 300 µg proteins of RAW264.7 cells expressing TurboID-STING were compared by LC-MS/MS analysis. We assessed reproducibility of each workflow using six technical replicates. Original workflo...
ORGANISM(S): Mus Musculus (mouse) 
2022-08-25 | PXD035220 | 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
ProteomicsPRM of DMXAA-treated Raw264.7 secretome
Raw264.7 cells were cultured in serum-free DMEM containing DMSO or DMXAA for 4 h, and the conditioned media were collected. The precipitated proteins were directly digested with trypsin/Lys-C mix. Three biological replicates of the samples were individually prepared and analyzed by targeted LC-MS/MS...
ORGANISM(S): Mus Musculus (mouse) 
2018-04-04 | PXD009170 | JPOST Repository
ProteomicsLFQ of DMXAA-treated Raw264.7 secretome
Raw264.7 cells were cultured in serum-free DMEM containing DMSO or DMXAA for 4 h, and the conditioned medium were collected. The precipitated proteins were directly digested with trypsin. Three biological replicates of the samples were individually prepared and analyzed by LC-MS/MS. Label-free quant...
ORGANISM(S): Mus Musculus (mouse) 
2018-04-04 | PXD009169 | JPOST Repository
Proteomics50_100uM_DSBU_XL-MS_PHB2(V1)-Myc_HEK293
Mitochondria were isolated from HEK293 cells that stably express PHB2(V1) with an C-terminal 3×Myc tag, and were resuspended in a cross-linking buffer [1 × PBS (pH 7.2) and 50 or 100 μM disuccinimidyl didutyric urea (DSBU) for 30 min at room temperature. The cross-linked mitochondria were then lysed...
ORGANISM(S): Homo Sapiens (human) 
2019-09-05 | PXD011941 | JPOST Repository
Proteomics100uM_DSBU_XL-MS_PHB1-Myc_HEK293
Mitochondria were isolated from HEK293 cells that stably express PHB1 with an C-terminal 3×Myc tag, and were resuspended in a cross-linking buffer [1 × PBS (pH 7.2) and 100 μM disuccinimidyl didutyric urea (DSBU) for 30 min at room temperature. The cross-linked mitochondria were then lysed in RIPA b...
ORGANISM(S): Homo Sapiens (human) 
2019-09-05 | PXD014217 | JPOST Repository
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