rabbit polyclonal anti pdk1  (Proteintech)

Journal: Cell Reports

Article Title: HIF1α-dependent uncoupling of glycolysis suppresses tumor cell proliferation

doi: 10.1016/j.celrep.2024.114103

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Article Snippet: Rabbit polyclonal anti- PDK1 , Enzo Life Sciences, Inc. , Cat#ADI-KAP-PK112; RRID: AB_10618932.

Techniques: Transduction, Recombinant, DC Protein Assay, Bicinchoninic Acid Protein Assay, Western Blot, Modification, Transfection, Mass Spectrometry

Journal: Particle and Fibre Toxicology

Article Title: Titanium dioxide and carbon black nanoparticles disrupt neuronal homeostasis via excessive activation of cellular prion protein signaling

doi: 10.1186/s12989-022-00490-x

Figure Lengend Snippet: TiO 2 and CB nanoparticle interaction with PrP C promotes TACE internalization in a PDK1-dependent manner at the root of TNFR1 overexposure at the cell surface. a TACE immunostaining at the plasma membrane of 1C11 cells exposed to TiO 2 - or CB-NPs (1 µg cm −2 ) for 1 h in the presence or not of a siRNA toward PrP C (siPrP) or the PDK1 inhibitor, BX912 (1 µM) and related quantification histogram. Cell permeabilization with saponin (0.05%) shows TACE internalization in 1C11 cells exposed to nanoparticles. Scale bar = 10 µm. b , c TACE expression level as assessed by RT-qPCR b and Western-blotting c in 1C11 cells exposed to TiO 2 - or CB-NPs (1 µg cm −2 ) for 1 h. d PDK1 phosphorylation status at Ser241 (p-PDK1) was assessed by Western-blotting in 1C11 cells exposed to TiO 2 - or CB-NPs (1 µg cm −2 ) for 1 h. e TNFR1 immunostaining at the plasma membrane of 1C11 cells exposed for 1 h to TiO 2 - or CB-NPs (1 µg cm −2 ) in the presence or not of a siRNA toward PrP C (siPrP) or the PDK1 inhibitor, BX912 (1 µM) and related quantification histogram. Scale bar = 10 µm. The experiments were performed three times in triplicates. Values are means ± SEM. *denotes p < 0.05, ** p < 0.01, *** p < 0.001 versus unexposed cells

Article Snippet: Rabbit polyclonal PDK1 and Ser241 phospho-specific PDK1 antibodies were purchased from Cell Signaling (Leiden, The Netherlands) to measure PDK1 level and indirectly PDK1 activity by Western-blot.

Techniques: Immunostaining, Clinical Proteomics, Membrane, Expressing, Quantitative RT-PCR, Western Blot, Phospho-proteomics

Journal: Particle and Fibre Toxicology

Article Title: Titanium dioxide and carbon black nanoparticles disrupt neuronal homeostasis via excessive activation of cellular prion protein signaling

doi: 10.1186/s12989-022-00490-x

Figure Lengend Snippet: Cell exposure to TiO 2 and CB nanoparticles promotes rise of Aβ42 through corruption of the PrP C -PDK1 pathway. a Kinetics of intracellular Aβ accumulation in 1C11 cells exposed to TiO 2 - or CB-NPs (1 µg cm −2 ) and related quantification histogram of immunostained Aβ. Scale bar = 10 µm. b ELISA-based quantification of Aβ42 peptides in 1C11 and 1C11 5−HT neuronal cells exposed to TiO 2 - or CB-NPs (1 µg cm −2 ) up to 24 h. c ELISA-based quantification of Aβ42 peptides in 1C11 5−HT neuronal cells exposed to TiO 2 - or CB-NPs (1 µg cm −2 ) for 4 h in the presence or not of a siRNA toward PrP C (siPrP) or the PDK1 inhibitor, BX912 (1 µM). The experiments were performed three times in triplicates. Values are means ± SEM. *denotes p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus unexposed cells

Article Snippet: Rabbit polyclonal PDK1 and Ser241 phospho-specific PDK1 antibodies were purchased from Cell Signaling (Leiden, The Netherlands) to measure PDK1 level and indirectly PDK1 activity by Western-blot.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Journal of Exercise Rehabilitation

Article Title: Effect of treadmill exercise and bone marrow stromal cell engraftment on activation of BDNF-ERK-CREB signaling pathway in the crushed sciatic nerve

doi: 10.12965/jer.2142626.313

Figure Lengend Snippet: Combination of exercise training and bone marrow stromal cell (BMSC) engraftment activated BDNF-Akt signaling pathway 28 days after sciatic nerve injury. Only EX+TP group strongly increased BDNF levels, but p-PDK1 was significantly enhanced in EX, TP, and EX+TP groups than those in NEX group. Additionally, EX+TP group leads to greater phosphorylation of Akt compared to EX group. Upper panel: representative expression of BDNF, phosphorylated PDK-1 (p-PDK-1), and p-Akt. Lower left panel: the density ratio of BDNF to β-actin band. Lower middle panel: the density ratio of p-PDK-1 to β-actin band. Lower right panel: the density ratio of p-Akt to β-actin band. BDNF, brain-derived neurotrophic factor; Akt, protein kinase B; PDK-1, phosphoinositide-dependent kinase-1; NORM, normal group; NEX, nonexercise training group; EX, exercise training group; TP, BMSC transplantation group; EX+TP, exercise training, and BMSC transplantation group. * P <0.05, ** P <0.01, *** P <0.001 vs. SED group. ††† P <0.001 vs. EX group.

Article Snippet: Protein (20 μg) was used for Western blot analysis using anti-BDNF rabbit monoclonal antibody (1:1,000, Cell Signaling Biotechnology, Danvers, MA, USA), anti-phosphorylated 3-phosphoinositide-dependent protein kinase-1 (p-PDK1) rabbit polyclonal antibody (1:1,000, Cell Signaling Biotechnology), anti-phosphorylated protein kinase B (p-Akt) rabbit monoclonal antibody (1:1,000, Cell Signaling Biotechnology), anti-phosphorylated CREB rabbit monoclonal antibody (1:1,000, Cell Signaling Biotechnology), anti-ATF3 rabbit monoclonal antibody (1:1,000, Cell Signaling Biotechnology), anti-phosphorylated ERK1/2 (p-ERK1/2) rabbit monoclonal antibody (1:1,000, Cell Signaling Biotechnology), anti-β-actin (1:2,000, Santa Cruz Biotechnology, Santa Cruz, CA, USA) antibodies, and goat anti-mouse or goat anti-rabbit horseradish peroxidase-conjugated secondary antibody (1:1,000, GeneTex Inc., Irvine, CA, USA) were used.

Techniques: Phospho-proteomics, Expressing, Derivative Assay, Transplantation Assay

Journal: Cell reports

Article Title: Impact of WIN site inhibitor on the WDR5 interactome

doi: 10.1016/j.celrep.2020.108636

Figure Lengend Snippet: (A) Comparison of C6nc/C6 ratios for the two SILAC replicates. Depleted proteins that met a 2-fold cutoff in both replicates are highlighted in blue. (B) As in (A) except for enriched proteins (red). (C) Extracts from HEK293 cells were subject to IP with a polyclonal antibody against WDR5 or an immunoglobulin G (IgG) control. IP samples were probed with antibodies against the indicated endogenous proteins. Inputs are 2% for WDR5 and RBBP5 and 0.3% for others. n = 3 biological replicates. (D) As in (C) but for different candidate proteins. Inputs are 5% for WDR5, RBBP5, and UBR5 and 0.3% for others. n = 3 biological replicates. (E) HEK293 cells stably expressing FLAG-tagged WDR5 were treated for 4 h with 30 μM C6 or C6nc prior to lysis and subsequent FLAG IP. For ethidium bromide (EtBr) treatment, 200 μg/mL EtBr was added to the lysate for the duration of the experiment. Candidate WDR5 interaction partners were probed by IB. Inputs are 5% for WDR5 and RBBP5, 0.1% for SYRC and SYIC, and 1% for all others; n = 3 biological replicates. (F) HEK293 cells stably expressing FLAG-tagged WDR5 were treated for 4 h with 30 mM C6 or C6nc prior to lysis and subsequent FLAG IP in buffer using CHAPS detergent. IP samples were probed with antibodies against the indicated proteins. Inputs are 10% for WDR5 and RBBP5 and 1% for others; n = 3 biological replicates. (G) HEK293 cells stably expressing FLAG-tagged WDR5 proteins were treated for 4 h with 30 μM C6 (where indicated) prior to lysis and FLAG IP. IP samples were probed with antibodies against the indicated proteins. Inputs are 10% for WDR5 and 1% for others; n = 3 biological replicates. See also .

Article Snippet: Rabbit polyclonal anti-PDK1 antibody (used for IP and western blotting) , Bethyl Laboratories , Cat# A302–130A; RRID:AB_1720395.

Techniques: Comparison, Multiplex sample analysis, Control, Stable Transfection, Expressing, Lysis

Journal: Cell reports

Article Title: Impact of WIN site inhibitor on the WDR5 interactome

doi: 10.1016/j.celrep.2020.108636

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-PDK1 antibody (used for IP and western blotting) , Bethyl Laboratories , Cat# A302–130A; RRID:AB_1720395.

Techniques: Western Blot, Virus, Recombinant, Protease Inhibitor, Multiplex sample analysis, Staining, Cloning, Expressing, Transfection, Mass Spectrometry, Plasmid Preparation, Mutagenesis, CRISPR, Amplification, Modification, Software, Hybridization, Membrane