nos1 rabbit polyclonal antibody  (Santa Cruz Biotechnology)

Journal: PLoS ONE

Article Title: The Spinal Cord Expression of Neuronal and Inducible Nitric Oxide Synthases and Their Contribution in the Maintenance of Neuropathic Pain in Mice

doi: 10.1371/journal.pone.0014321

Figure Lengend Snippet: Development of mechanical allodynia ( A ), thermal hyperalgesia ( B ) and thermal allodynia ( C ) in the ipsilateral paw of WT, NOS1-KO and NOS2-KO mice at 0, 1, 4, 7, 10, 14 and 21 days after sciatic nerve ligation. For each test and time tested, * indicates significant differences when compared sciatic nerve-injured WT mice vs. sham-operated WT mice (* p<0.05, ** p<0.01, *** p<0.001, one-way ANOVA followed by Scheffe test), + when compared sciatic nerve-injured NOS1-KO mice vs. sciatic nerve-injured WT mice (+ p<0.05, ++ p<0.01, +++ p<0.001, one-way ANOVA followed by Scheffe test) and # when compared sciatic nerve-injured NOS2-KO mice vs. sciatic nerve-injured WT mice (# p<0.05, ## p<0.01, ### p<0.001, one-way ANOVA followed by Scheffe test). In the cold plate test (C), • indicates significant differences when compared sciatic nerve-injured NOS1-KO mice vs. sham-operated NOS1-KO mice (• p<0.05, •• p<0.01, one-way ANOVA followed by Scheffe test). Results are shown as mean values ± SEM; n = 10–12 animals per experimental group.

Article Snippet: The proteins were electrophoretically transferred onto PVDF membrane overnight, blocked with PBST +2% nonfat dry milk, and subsequently incubated overnight at 4°C with a polyclonal rabbit anti-NOS1 antibody (1∶150, BD Transduction Laboratories, San Diego, CA, USA), a polyclonal rabbit anti-NOS2 antibody (1∶200, Chemicon, Millipore), a polyclonal rabbit anti-NOS3 antibody (1∶100, BD Transduction Laboratories, San Diego, CA, USA), or a monoclonal rabbit anti-β-actin antibody (1∶10.000, Sigma, St. Louis, MO, USA). β-actin was used as a loading control.

Techniques: Ligation, Mouse Assay

Journal: PLoS ONE

Article Title: The Spinal Cord Expression of Neuronal and Inducible Nitric Oxide Synthases and Their Contribution in the Maintenance of Neuropathic Pain in Mice

doi: 10.1371/journal.pone.0014321

Figure Lengend Snippet: The relative NOS1 mRNA ( A ) and protein ( B ) expression in the ipsilateral site of the lumbar section of the spinal cord from sham-operated (SH) and sciatic nerve-injured (SNI) WT and NOS2-KO mice at 21 days after sciatic nerve ligation are represented. In both figures, * indicates significant differences when compared sciatic nerve-injured WT mice vs. sham-operated WT animals (* p<0.05, one-way ANOVA followed by Student-Newman-Keuls test). A representative example of Western blots for NOS1 protein (155 kDa) in which β-actin (43 kDa) was used as a loading control is shown in C . Data are expressed as mean values ± SEM; n = 4–5 samples per group.

Article Snippet: The proteins were electrophoretically transferred onto PVDF membrane overnight, blocked with PBST +2% nonfat dry milk, and subsequently incubated overnight at 4°C with a polyclonal rabbit anti-NOS1 antibody (1∶150, BD Transduction Laboratories, San Diego, CA, USA), a polyclonal rabbit anti-NOS2 antibody (1∶200, Chemicon, Millipore), a polyclonal rabbit anti-NOS3 antibody (1∶100, BD Transduction Laboratories, San Diego, CA, USA), or a monoclonal rabbit anti-β-actin antibody (1∶10.000, Sigma, St. Louis, MO, USA). β-actin was used as a loading control.

Techniques: Expressing, Ligation, Western Blot

Journal: PLoS ONE

Article Title: The Spinal Cord Expression of Neuronal and Inducible Nitric Oxide Synthases and Their Contribution in the Maintenance of Neuropathic Pain in Mice

doi: 10.1371/journal.pone.0014321

Figure Lengend Snippet: The relative NOS2 mRNA ( A ) and protein ( B ) expression in the ipsilateral site of the lumbar section of the spinal cord from sham-operated (SH) and sciatic nerve-injured (SNI) WT and NOS1-KO mice at 21 days after sciatic nerve ligation are represented. In both figures, * indicates significant differences when compared sciatic nerve-injured NOS1-KO mice vs. sham-operated NOS1-KO animals (* p<0.05, one-way ANOVA followed by Student-Newman-Keuls test). A representative example of Western blots for NOS2 protein (130 kDa) in which β-actin (43 kDa) was used as a loading control is shown in C . Data are expressed as mean values ± SEM; n = 4–5 samples per group.

Article Snippet: The proteins were electrophoretically transferred onto PVDF membrane overnight, blocked with PBST +2% nonfat dry milk, and subsequently incubated overnight at 4°C with a polyclonal rabbit anti-NOS1 antibody (1∶150, BD Transduction Laboratories, San Diego, CA, USA), a polyclonal rabbit anti-NOS2 antibody (1∶200, Chemicon, Millipore), a polyclonal rabbit anti-NOS3 antibody (1∶100, BD Transduction Laboratories, San Diego, CA, USA), or a monoclonal rabbit anti-β-actin antibody (1∶10.000, Sigma, St. Louis, MO, USA). β-actin was used as a loading control.

Techniques: Expressing, Ligation, Western Blot

Journal: PLoS ONE

Article Title: The Spinal Cord Expression of Neuronal and Inducible Nitric Oxide Synthases and Their Contribution in the Maintenance of Neuropathic Pain in Mice

doi: 10.1371/journal.pone.0014321

Figure Lengend Snippet: The relative NOS3 mRNA ( A ) and protein ( B ) expression in the ipsilateral site of the lumbar section of the spinal cord from sham-operated (SH) and sciatic nerve-injured (SNI) WT, NOS2-KO and NOS1-KO mice at 21 days after sciatic nerve ligation are represented. In both figures, * indicates significant differences when compared sham-operated NOS1-KO animals vs. the other groups. (* p<0.05, one-way ANOVA followed by Student-Newman-Keuls test). A representative example of Western blot for NOS3 protein (140 kDa) in which β-actin (43 kDa) was used as a loading control is shown in C . Data are expressed as mean values ± SEM; n = 4–5 samples per group.

Article Snippet: The proteins were electrophoretically transferred onto PVDF membrane overnight, blocked with PBST +2% nonfat dry milk, and subsequently incubated overnight at 4°C with a polyclonal rabbit anti-NOS1 antibody (1∶150, BD Transduction Laboratories, San Diego, CA, USA), a polyclonal rabbit anti-NOS2 antibody (1∶200, Chemicon, Millipore), a polyclonal rabbit anti-NOS3 antibody (1∶100, BD Transduction Laboratories, San Diego, CA, USA), or a monoclonal rabbit anti-β-actin antibody (1∶10.000, Sigma, St. Louis, MO, USA). β-actin was used as a loading control.

Techniques: Expressing, Ligation, Western Blot

Journal: bioRxiv

Article Title: Calcium transients in nNOS neurons underlie distinct phases of the neurovascular response to barrel cortex activation in awake mice

doi: 10.1101/2022.10.03.510654

Figure Lengend Snippet: A : GCaMP7f was expressed in NOS1 cre mice using systemic administration of an AAV-PHP.eB viral vector. The localization of GCaMP7f expression in nNOS neurons in the cortical layers was verified by immunohistochemistry (IHC). Right panels are enlargements of the dashed squares in the left panel (n=2,744 neurons in 5 mice). B : GCaMP7f expression in inhibitory (GAD67) or excitatory (CaMKII) neurons of nNOS cre mice treated with the AAV-PHP.eB viral vector. C : Neurons (%) expressing nNOS, GCaMP7f or both, and, among GCaMP7f positive neurons, those expressing GAD67 or CaMKII (n=690 neurons in 3 mice). D : Single-cell RNAseq data (mousebrain.org/development/downloads.html) showing expression of Nos1 both in excitatory ( Slc17a7 ) and inhibitory ( Gad1 ) neurons (%), confirming the finding by immunohistochemistry ( B ). Scale bar=50μm throughout.

Article Snippet: The following primary antibodies were used: rabbit anti-NOS1 polyclonal antibody (1:300, AB5380, Sigma-Aldrich), chicken anti-GFP polyclonal antibody (1:500, A10262, Invitrogen), mouse anti-GAD67 monoclonal antibody (1:300, MAB5406, Sigma-Aldrich), rabbit anti-CaMKII monoclonal antibody (1:300, ab52476, abcam).

Techniques: Plasmid Preparation, Expressing, Immunohistochemistry

Journal: Neuron

Article Title: Temporally and Spatially Distinct Thirst Satiation Signals

doi: 10.1016/j.neuron.2019.04.039

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-Nos1 , Santa Cruz , Cat#sc-648; RRID: AB_630935.

Techniques: Virus, Plasmid Preparation, Injection, Recombinant, Knock-In, Software

Journal: Molecules

Article Title: The Up-Regulation of Oxidative Stress as a Potential Mechanism of Novel MAO-B Inhibitors for Glioblastoma Treatment

doi: 10.3390/molecules24102005

Figure Lengend Snippet: Western blotting analysis of NOS-1, NOS-2 and VEGF expression in rat C6 glioma cell lines treated with Cmp 5 and Cmp 3 . ( A ) Cells treated with DMSO (0.2%) were loaded as the negative control. Each membrane has been probed with β–actin antibody to verify loading consistency. Western blot is the most representative of three different experiments. ( B – D ) Histograms represent densitometric measurements of proteins bands expressed as integrated optical intensity (IOI) mean of three separate experiments. The error bars on these graphs show standard deviation (± SD). Densitometric values analysed by ANOVA (post hoc application of Tukey’s multiple comparison test) return significant differences. **** p < 0.0001, *** p < 0.0002, ** p < 0.0005, * p < 0.005.

Article Snippet: Nitrocellulose membranes, blocked in 5% of non-fat milk in PBS 0.1% Tween-20, were probed with mouse monoclonal anti-β actin antibody (antibody dilution 1:5000) (A5316 Sigma, MO, USA), rabbit polyclonal anti-VEGF anti-NOS-2 antibodies (antibodies dilution 1:200) (sc-152, sc-651, respectively, both purchased by Santa Cruz biotechnology, CA, USA), mouse monoclonal anti-HIF-1α, anti-MMP-9, anti-MMP-2, anti-NOS-1 antibodies (antibodies dilution 1:200) (sc-5354, sc-393859, sc-13595, sc-5302, respectively, all purchased by Santa Cruz biotechnology, CA, USA), then incubated in the presence of specific enzyme conjugated IgG horseradish peroxidase.

Techniques: Western Blot, Expressing, Negative Control, Standard Deviation