Incubation:Article Title: Fetal Neuronal Vesicles in the Assessment of Perinatal Brain Dysfunction and Late-Onset Growth Restriction: A Pilot Study
Article Snippet: .. The membranes were blocked with 5% NFDM/TBST for 2 h. Furthermore, incubation during an hour at the room temperature with primary antibodies, including BDNF (1:1000, ab205067, Abcam, Waltham, MA, USA), NGF (1:1000, SI79-01 (rabbit), Thermo Fisher Scientific, Waltham, MA, USA), NT3 (1:200, sc-518099, Santa Cruz Biotechnology, Santa Cruz, CA, USA), NT4 (1:200, sc-365444, Santa Cruz Biotechnology, Santa Cruz, CA, USA), SYNAPTOTAGMIN 1 (SYT1) (1:200, sc-136480, Santa Cruz Biotechnology, Santa Cruz, CA, USA), SYNAPSIN 1 (SYN1) (1:1000, ab254349 (rabbit), Abcam, Waltham, MA, USA), SYNAPTOPHYSIN (SYP) (1:1000, sc-17750, Santa Cruz Biotechnology, Santa Cruz, CA, USA), SYNAPTOPODIN (SYNPO) (1:100, sc-515842, Santa Cruz Biotechnology, Santa Cruz, CA, USA), PSD95 (1:200, sc-32291, Santa Cruz Biotechnology, Santa Cruz, CA, USA), Neuronal Pentraxin 2 (NPTX2) (1:1000, ab277523 (rabbit), Abcam, Waltham, MA, USA), NEDD8 (1:1000, ab81264, (rabbit), Abcam, Waltham, MA, USA), NAE1 (APPBP1) (1:100, sc-390002, Santa Cruz Biotechnology, Santa Cruz, CA, USA), UBC12 (1:100, sc-390064, Santa Cruz Biotechnology, Santa Cruz, CA, USA), SUMO 1 (1:200; sc-5308, Santa Cruz Biotechnology, Santa Cruz, CA, USA), SUMO 2/3/4 (1:200, sc-393144, Santa Cruz Biotechnology, Santa Cruz, CA, USA), UBC9 (1:100; sc-271057, Santa Cruz Biotechnology, Santa Cruz, CA, USA), Contactin-2/TAG1 (1:1000; MAB17141, Fisher Scientific, Waltham, MA, USA). .. Exosomal marker CD81 was used to assess plasma FNVs levels (1:100, sc-166029, Santa Cruz Biotechnology, Santa Cruz, CA, USA).
Article Title: Fetal Neuronal Vesicles in the Assessment of Perinatal Brain Dysfunction and Late-Onset Growth Restriction: A Pilot Study.
Article Snippet: .. The membranes were blocked with 5% NFDM/TBST for 2 h. Furthermore, incubation during an hour at the room temperature with primary antibodies, including BDNF (1:1000, ab205067, Abcam, Waltham, MA, USA), NGF (1:1000, SI79-01 (rabbit), Thermo Fisher Scientific, Waltham, MA, USA), NT3 (1:200, sc-518099, Santa Cruz Biotechnology, Santa Cruz, CA, USA), NT4 (1:200, sc-365444, Santa Cruz Biotechnology, Santa Cruz, CA, USA), SYNAPTOTAGMIN 1 (SYT1) (1:200, sc-136480, Santa Cruz Biotechnology, Santa Cruz, CA, USA), SYNAPSIN 1 (SYN1) (1:1000, ab254349 (rabbit), Abcam, Waltham, MA, USA), SYNAPTOPHYSIN (SYP) (1:1000, sc-17750, Santa Cruz Biotechnology, Santa Cruz, CA, USA), SYNAPTOPODIN (SYNPO) (1:100, sc-515842, Santa Cruz Biotechnology, Santa Cruz, CA, USA), PSD95 (1:200, sc-32291, Santa Cruz Biotechnology, Santa Cruz, CA, USA), Neuronal Pentraxin 2 (NPTX2) (1:1000, ab277523 (rabbit), Abcam, Waltham, MA, USA), NEDD8 (1:1000, ab81264, (rabbit), Abcam, Waltham, MA, USA), NAE1 (APPBP1) (1:100, sc-390002, Santa Cruz Biotechnology, Santa Cruz, CA, USA), UBC12 (1:100, sc-390064, Santa Cruz Biotechnology, Santa Cruz, CA, USA), SUMO 1 (1:200; sc-5308, Santa Cruz Biotechnology, Santa Cruz, CA, USA), SUMO 2/3/4 (1:200, sc-393144, Santa Cruz Biotechnology, Santa Cruz, CA, USA), UBC9 (1:100; sc-271057, Santa Cruz Biotechnology, Santa Cruz, CA, USA), Contactin-2/TAG1 (1:1000; MAB17141, Fisher Scientific, Waltham, MA, USA). .. Exosomal marker CD81 was used to assess plasma FNVs levels (1:100, sc-166029, Santa Cruz Biotechnology, Santa Cruz, CA, USA).
Article Title: Conditioned medium from human gingival mesenchymal stem cells protects motor-neuron-like NSC-34 cells against scratch-injury-induced cell death.
Article Snippet: After blotting, the PVDF membranes were stained with Ponceau S solution (SERVA, Denmark) to assess protein separation. .. Then, the membranes were incubated in blocking solution (5% skimmed milk in 1× PBS) at ambient temperature for 1 h. After blocking, the membranes were incubated with the following antibodies for overnight at 4°C: nerve growth factor (NGF) (1:250; Abcam, USA), NT3 (1:250; Santa Cruz Biotechnology Inc., USA), IL-10 (1:250; Santa Cruz Biotechnology Inc., USA), and transforming growth factor beta (TGF-β) (1:500; Abcam, USA). .. Subsequently, membranes were incubated with appropriate HRP-conjugated secondary antibodies IgG (1:2000; Santa Cruz Biotechnology Inc., USA).
Planar Chromatography:Article Title: Stress Diminishes BDNF-stimulated TrkB Signaling, TrkB-NMDA Receptor Linkage and Neuronal Activity in the Rat Brain.
Article Snippet: Anti-PSD-95 (05494) was from Millipore. .. AntiTrkB (SC-8316), -pTrkB (SC-8058), -BDNF/pro-BDNF (SC-2098), -NT3 (SC-547), -NT4 (SC-545), - phosphotyrosine (SC-508), -Erk2 (SC-154, SC-81457), - pErk2 (SC-7383), -pAkt1 (SC-7985-R), -Akt1 (SC65487), -PLC (SC-7290), -NR1 (SC-9058), -NR2A (SC9056), -actin (SC-376421), -b-actin (SC-47778), and - Arc (SC-365736) were from Santa Cruz Biotechnology. .. Seize-X immunoprecipitation kit, antigen elution buffer, Bind NeutrAvidin, high binding capacity coated 96-well plates, and West Pico chemiluminescent reagents were from Pierce-Endogen.
Isolation:Article Title: Neurotrophin NT3 promotes ovarian primordial to primary follicle transition
Article Snippet: .. Ovary sections from freshly isolated adult or cultured postnatal day 4 (P4) ovaries (unrelated control ovaries cultured for 10 days) were immunostained as previously described ( Nilsson et al . 2002 ) for the presence of NT3 using anti-NT3 antibody (Anti-NT3 rabbit IgG, 1–0.1 μg/ml; Santa Cruz Biotechnology, Santa Cruz, CA, USA), or for NTRK3 using anti-NTRK3 antibody (Anti-NTRK3 rabbit IgG, 1–10 μg/ml; Santa Cruz Biotechnology). ..
Cell Culture:Article Title: Neurotrophin NT3 promotes ovarian primordial to primary follicle transition
Article Snippet: .. Ovary sections from freshly isolated adult or cultured postnatal day 4 (P4) ovaries (unrelated control ovaries cultured for 10 days) were immunostained as previously described ( Nilsson et al . 2002 ) for the presence of NT3 using anti-NT3 antibody (Anti-NT3 rabbit IgG, 1–0.1 μg/ml; Santa Cruz Biotechnology, Santa Cruz, CA, USA), or for NTRK3 using anti-NTRK3 antibody (Anti-NTRK3 rabbit IgG, 1–10 μg/ml; Santa Cruz Biotechnology). ..
Control:Article Title: Neurotrophin NT3 promotes ovarian primordial to primary follicle transition
Article Snippet: .. Ovary sections from freshly isolated adult or cultured postnatal day 4 (P4) ovaries (unrelated control ovaries cultured for 10 days) were immunostained as previously described ( Nilsson et al . 2002 ) for the presence of NT3 using anti-NT3 antibody (Anti-NT3 rabbit IgG, 1–0.1 μg/ml; Santa Cruz Biotechnology, Santa Cruz, CA, USA), or for NTRK3 using anti-NTRK3 antibody (Anti-NTRK3 rabbit IgG, 1–10 μg/ml; Santa Cruz Biotechnology). ..
Blocking Assay:Article Title: Conditioned medium from human gingival mesenchymal stem cells protects motor-neuron-like NSC-34 cells against scratch-injury-induced cell death.
Article Snippet: After blotting, the PVDF membranes were stained with Ponceau S solution (SERVA, Denmark) to assess protein separation. .. Then, the membranes were incubated in blocking solution (5% skimmed milk in 1× PBS) at ambient temperature for 1 h. After blocking, the membranes were incubated with the following antibodies for overnight at 4°C: nerve growth factor (NGF) (1:250; Abcam, USA), NT3 (1:250; Santa Cruz Biotechnology Inc., USA), IL-10 (1:250; Santa Cruz Biotechnology Inc., USA), and transforming growth factor beta (TGF-β) (1:500; Abcam, USA). .. Subsequently, membranes were incubated with appropriate HRP-conjugated secondary antibodies IgG (1:2000; Santa Cruz Biotechnology Inc., USA).
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