Western Blot:Article Title: Primary microRNA precursor transcripts are localized at post-synaptic densities in adult mouse forebrain.
Article Snippet: Primer specificity was verified by a single peak in the melting-curve analysis and a single band of the right size on a 2.5% agarose gel (NuSieve 3 : 1; Lonza, Basel, Switzerland) for all genes analyzed. .. Western blotting was performed as described (Lugli et al. 2005, 2008). polyvinylidene difluoride blots were blocked in 1% non-fat dry milk for 1 h, room temperature, incubated with primary antiPSD95 rabbit polyclonal antibody (1 : 30 000, D74D3 Cell Signaling Technology, Danvers, MA, USA), anti-Drosha antibody (1 : 2500, raised in chicken by us against a peptide sequence located at the C-terminus RWEREHQEREPEEAEDIKK), anti-Drosha polyclonal antibody (1 : 2000, SAB2500885, raised in goat; Sigma-Aldrich, St. Louis, MO, USA), anti-DGCR8 monoclonal antibody (1 : 5000, 60084-1; Proteintech, Chicago, IL, USA), antispinophilin rabbit polyclonal antibody (1 : 5000, Sigma N-5162), anti-Kinesin KIF5 heavy-chain mouse monoclonal antibody (1 : 1000, MAB1614; EMD Millipore, Billerica, MA, USA), Kinesin light-chain mouse monoclonal antibody (1 : 1000, generous gift from Dr. Scott Brady), anti-dicer rabbit polyclonal antibody (1 : 10 000, described in Lugli et al. 2005), anti-Ago rabbit polyclonal antibody (1 : 10 000, described in Lugli et al. 2005), or anti-Staufen rabbit polyclonal antibody (1 : 2000, MB8277; Chemicon EMD Millipore, Billerica, MA, USA) overnight with Journal of Neurochemistry © 2012 International Society for Neurochemistry, J. Neurochem. (2012) 123, 459--466 © 2012 The Authors rocking at 4°C and rinsed. .. Goat anti-rabbit IgG (peroxidase conjugated; Sigma A0545), sheep anti-mouse IgG (peroxidase conjugates; GE Healthcare NA931), or goat anti-chicken IgY (peroxidase conjugated; Sigma A9046) were added at 1 : 30 000– 1 : 50 000 for 2 h and rinsed; finally, blots were incubated in ECLPlus reagent (Amersham GE Healthcare) and exposed to film (Hyperfilm ECL; Amersham).
Incubation:Article Title: Primary microRNA precursor transcripts are localized at post-synaptic densities in adult mouse forebrain.
Article Snippet: Primer specificity was verified by a single peak in the melting-curve analysis and a single band of the right size on a 2.5% agarose gel (NuSieve 3 : 1; Lonza, Basel, Switzerland) for all genes analyzed. .. Western blotting was performed as described (Lugli et al. 2005, 2008). polyvinylidene difluoride blots were blocked in 1% non-fat dry milk for 1 h, room temperature, incubated with primary antiPSD95 rabbit polyclonal antibody (1 : 30 000, D74D3 Cell Signaling Technology, Danvers, MA, USA), anti-Drosha antibody (1 : 2500, raised in chicken by us against a peptide sequence located at the C-terminus RWEREHQEREPEEAEDIKK), anti-Drosha polyclonal antibody (1 : 2000, SAB2500885, raised in goat; Sigma-Aldrich, St. Louis, MO, USA), anti-DGCR8 monoclonal antibody (1 : 5000, 60084-1; Proteintech, Chicago, IL, USA), antispinophilin rabbit polyclonal antibody (1 : 5000, Sigma N-5162), anti-Kinesin KIF5 heavy-chain mouse monoclonal antibody (1 : 1000, MAB1614; EMD Millipore, Billerica, MA, USA), Kinesin light-chain mouse monoclonal antibody (1 : 1000, generous gift from Dr. Scott Brady), anti-dicer rabbit polyclonal antibody (1 : 10 000, described in Lugli et al. 2005), anti-Ago rabbit polyclonal antibody (1 : 10 000, described in Lugli et al. 2005), or anti-Staufen rabbit polyclonal antibody (1 : 2000, MB8277; Chemicon EMD Millipore, Billerica, MA, USA) overnight with Journal of Neurochemistry © 2012 International Society for Neurochemistry, J. Neurochem. (2012) 123, 459--466 © 2012 The Authors rocking at 4°C and rinsed. .. Goat anti-rabbit IgG (peroxidase conjugated; Sigma A0545), sheep anti-mouse IgG (peroxidase conjugates; GE Healthcare NA931), or goat anti-chicken IgY (peroxidase conjugated; Sigma A9046) were added at 1 : 30 000– 1 : 50 000 for 2 h and rinsed; finally, blots were incubated in ECLPlus reagent (Amersham GE Healthcare) and exposed to film (Hyperfilm ECL; Amersham).
Sequencing:Article Title: Primary microRNA precursor transcripts are localized at post-synaptic densities in adult mouse forebrain.
Article Snippet: Primer specificity was verified by a single peak in the melting-curve analysis and a single band of the right size on a 2.5% agarose gel (NuSieve 3 : 1; Lonza, Basel, Switzerland) for all genes analyzed. .. Western blotting was performed as described (Lugli et al. 2005, 2008). polyvinylidene difluoride blots were blocked in 1% non-fat dry milk for 1 h, room temperature, incubated with primary antiPSD95 rabbit polyclonal antibody (1 : 30 000, D74D3 Cell Signaling Technology, Danvers, MA, USA), anti-Drosha antibody (1 : 2500, raised in chicken by us against a peptide sequence located at the C-terminus RWEREHQEREPEEAEDIKK), anti-Drosha polyclonal antibody (1 : 2000, SAB2500885, raised in goat; Sigma-Aldrich, St. Louis, MO, USA), anti-DGCR8 monoclonal antibody (1 : 5000, 60084-1; Proteintech, Chicago, IL, USA), antispinophilin rabbit polyclonal antibody (1 : 5000, Sigma N-5162), anti-Kinesin KIF5 heavy-chain mouse monoclonal antibody (1 : 1000, MAB1614; EMD Millipore, Billerica, MA, USA), Kinesin light-chain mouse monoclonal antibody (1 : 1000, generous gift from Dr. Scott Brady), anti-dicer rabbit polyclonal antibody (1 : 10 000, described in Lugli et al. 2005), anti-Ago rabbit polyclonal antibody (1 : 10 000, described in Lugli et al. 2005), or anti-Staufen rabbit polyclonal antibody (1 : 2000, MB8277; Chemicon EMD Millipore, Billerica, MA, USA) overnight with Journal of Neurochemistry © 2012 International Society for Neurochemistry, J. Neurochem. (2012) 123, 459--466 © 2012 The Authors rocking at 4°C and rinsed. .. Goat anti-rabbit IgG (peroxidase conjugated; Sigma A0545), sheep anti-mouse IgG (peroxidase conjugates; GE Healthcare NA931), or goat anti-chicken IgY (peroxidase conjugated; Sigma A9046) were added at 1 : 30 000– 1 : 50 000 for 2 h and rinsed; finally, blots were incubated in ECLPlus reagent (Amersham GE Healthcare) and exposed to film (Hyperfilm ECL; Amersham).
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