mouse anti pacap  (Santa Cruz Biotechnology)

Journal: International Journal of Molecular Sciences

Article Title: Differences in Neuropathology between Nitroglycerin-Induced Mouse Models of Episodic and Chronic Migraine

doi: 10.3390/ijms25073706

Figure Lengend Snippet: Neuropeptide expression patterns in the Sp5C in mouse models of episodic and chronic NTG-induced migraine (EM and CM, respectively). ( A ) Representative images of VIP, CGRP, PACAP, and substance P (green) immunofluorescence staining in the Sp5C of EM and CM mouse models. In the merged results, DAPI was added along with the VIP, CGRP, PACAP, and substance P markers. Scale bars = 20 μm. ( B ) Fluorescence intensities of VIP, CGRP, PACAP, and substance P in the Sp5C of EM and CM mouse models. One-way ANOVA with post hoc Tukey test: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. ns, not significant.

Article Snippet: To examine neuron activation, brain tissue slices were permeabilized in 0.5% phosphate-buffered saline with Triton X-100 for 5 min, incubated in a blocking solution at room temperature (RT) for 1 h, and incubated with primary antibodies for double staining of c-Fos (Cat No. 226008, 1:200; Synaptic Systems, Göttingen, Germany) and NeuN (Cat No. ab104224, 1:200; Abcam, Cambridge, UK) at 4 °C for 16 h, then at RT for 1 h. To examine increases in microglial activation, staining of Iba1 (Cat No. ab22378, 1:200; Abcam) was performed at 4 °C for 16 h, then at RT for 1 h. To compare neurotransmitter changes in the trigeminal spinal nucleus caudalis (Sp5C) and anterior cingulate cortex (ACC) regions of the brain, the following markers were assessed via staining: vasoactive intestinal peptide (VIP) (Cat No. ab272726, 1:200; Abcam), calcitonin gene-related peptide (CGRP) (Cat No. ab81887, 1:200; Abcam), pituitary adenylate cyclase-activating polypeptide (PACAP) (Cat No. sc-166180, 1:200; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and substance P (Cat No. ab14184, 1:200; Abcam) at 4 °C for 16 h, then at RT for 1 h. After the slides had been washed, they were incubated with the appropriate Alexa-Fluor 488- or 594-conjugated secondary antibody (1:500) at RT for 1 h. For nuclear staining, slides were incubated with 4′,6-diamidino-2-phenylindole (DAPI, 1:10,000) for 20 min before mounting.

Techniques: Expressing, Immunofluorescence, Staining, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Differences in Neuropathology between Nitroglycerin-Induced Mouse Models of Episodic and Chronic Migraine

doi: 10.3390/ijms25073706

Figure Lengend Snippet: Neuropeptide expression patterns in the ACC in mouse models of episodic and chronic NTG-induced migraine (EM and CM, respectively). ( A ) Representative images of VIP, CGRP, PACAP, and substance P (green) immunofluorescence staining in the ACC of EM and CM mouse models. In the merged results, DAPI was added along with the VIP, CGRP, PACAP, and substance P markers. Scale bars = 20 μm. ( B ) Fluorescence intensities of VIP, CGRP, PACAP, and substance P in the ACC of EM and CM mouse models. Statistical analysis was conducted using the Kruskal–Wallis test with Dunn’s multiple comparisons test and one-way ANOVA with post hoc Tukey test: * p < 0.05; ** p < 0.01; **** p < 0.0001; ns, not significant. Abbreviations: ACC, anterior cingulate cortex; CGRP, calcitonin gene-related peptide; NTG, nitroglycerin; PACAP, pituitary adenylate cyclase-activating peptide; VEH, vehicle control; VIP, vasoactive intestinal peptide.

Article Snippet: To examine neuron activation, brain tissue slices were permeabilized in 0.5% phosphate-buffered saline with Triton X-100 for 5 min, incubated in a blocking solution at room temperature (RT) for 1 h, and incubated with primary antibodies for double staining of c-Fos (Cat No. 226008, 1:200; Synaptic Systems, Göttingen, Germany) and NeuN (Cat No. ab104224, 1:200; Abcam, Cambridge, UK) at 4 °C for 16 h, then at RT for 1 h. To examine increases in microglial activation, staining of Iba1 (Cat No. ab22378, 1:200; Abcam) was performed at 4 °C for 16 h, then at RT for 1 h. To compare neurotransmitter changes in the trigeminal spinal nucleus caudalis (Sp5C) and anterior cingulate cortex (ACC) regions of the brain, the following markers were assessed via staining: vasoactive intestinal peptide (VIP) (Cat No. ab272726, 1:200; Abcam), calcitonin gene-related peptide (CGRP) (Cat No. ab81887, 1:200; Abcam), pituitary adenylate cyclase-activating polypeptide (PACAP) (Cat No. sc-166180, 1:200; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and substance P (Cat No. ab14184, 1:200; Abcam) at 4 °C for 16 h, then at RT for 1 h. After the slides had been washed, they were incubated with the appropriate Alexa-Fluor 488- or 594-conjugated secondary antibody (1:500) at RT for 1 h. For nuclear staining, slides were incubated with 4′,6-diamidino-2-phenylindole (DAPI, 1:10,000) for 20 min before mounting.

Techniques: Expressing, Immunofluorescence, Staining, Fluorescence, Control

Journal: International Journal of Molecular Sciences

Article Title: Differences in Neuropathology between Nitroglycerin-Induced Mouse Models of Episodic and Chronic Migraine

doi: 10.3390/ijms25073706

Figure Lengend Snippet: Statistical analysis of neuropeptide expression patterns in the Sp5C and ACC in mouse models of episodic and chronic NTG-induced migraine (EM and CM, respectively), * p < 0.05; *** p < 0.001; **** p < 0.0001.

Article Snippet: To examine neuron activation, brain tissue slices were permeabilized in 0.5% phosphate-buffered saline with Triton X-100 for 5 min, incubated in a blocking solution at room temperature (RT) for 1 h, and incubated with primary antibodies for double staining of c-Fos (Cat No. 226008, 1:200; Synaptic Systems, Göttingen, Germany) and NeuN (Cat No. ab104224, 1:200; Abcam, Cambridge, UK) at 4 °C for 16 h, then at RT for 1 h. To examine increases in microglial activation, staining of Iba1 (Cat No. ab22378, 1:200; Abcam) was performed at 4 °C for 16 h, then at RT for 1 h. To compare neurotransmitter changes in the trigeminal spinal nucleus caudalis (Sp5C) and anterior cingulate cortex (ACC) regions of the brain, the following markers were assessed via staining: vasoactive intestinal peptide (VIP) (Cat No. ab272726, 1:200; Abcam), calcitonin gene-related peptide (CGRP) (Cat No. ab81887, 1:200; Abcam), pituitary adenylate cyclase-activating polypeptide (PACAP) (Cat No. sc-166180, 1:200; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and substance P (Cat No. ab14184, 1:200; Abcam) at 4 °C for 16 h, then at RT for 1 h. After the slides had been washed, they were incubated with the appropriate Alexa-Fluor 488- or 594-conjugated secondary antibody (1:500) at RT for 1 h. For nuclear staining, slides were incubated with 4′,6-diamidino-2-phenylindole (DAPI, 1:10,000) for 20 min before mounting.

Techniques: Expressing