polyclonal nav 1 8 primary antibody  (Alomone Labs)

Journal: Journal of Neuroinflammation

Article Title: Functional up-regulation of Na v 1.8 sodium channel in Aβ afferent fibers subjected to chronic peripheral inflammation

doi: 10.1186/1742-2094-11-45

Figure Lengend Snippet: Cellular distribution of Na v 1.8 in rat DRG neurons. (A–F) Immunohistochemical distribution of Na v 1.8 in DRG neurons isolated from sham and CFA-treated rats. Panels show the localization of Na v 1.8 (green) in NF200-positive neurons (red, arrowheads). Yellow signal indicates double-labeled neurons. After inflammation, numerous large NF200-positive ganglion cells co-express Na v 1.8 (arrows). (G) Proportion of Na v 1.8-immunoreactive neurons in lumbar DRGs. The number of neurons expressing Na v 1.8 increases in CFA-inflamed rats compared to sham. (H) Percentage of NF200-containing DRG neurons expressing Na v 1.8. The proportion of dually stained cells increases after CFA-induced chronic inflammation. Data in panels G and H shown as mean ± SEM. Asterisks denote a statistically significant increase as compared with sham (4 rats/group, 10 sections/rat; *** P <0.001 or ** P <0.01; ANOVA followed by Sidak’s multiple comparisons test MCT)). # Statistically different from day 3; ### P <0.001, ## P <0.01, # P <0.05). Scale bars: 200 μm in A, C, and E and 20 μm in B, D, and F .

Article Snippet: To detect Na v 1.8, sections were incubated with the rabbit polyclonal anti-Na v 1.8 antibody (1:200; Alomone Labs, Jerusalem, Israel) in PBS containing 0.05% Triton X-100 and 0.5% NGS.

Techniques: Immunohistochemical staining, Isolation, Labeling, Expressing, Staining

Journal: Journal of Neuroinflammation

Article Title: Functional up-regulation of Na v 1.8 sodium channel in Aβ afferent fibers subjected to chronic peripheral inflammation

doi: 10.1186/1742-2094-11-45

Figure Lengend Snippet: Changes in the expression and distribution of Na v 1.8 associated with CFA-induced inflammation in rats. (A) Na v 1.8 mRNA levels of the ipsilateral hind paw are determined by qRT-PCR for sham animals and 14 days following CFA injection. Data are expressed as mean ± SEM (6–8 rats/group). ** P <0.01, CFA alone vs. sham (unpaired Student’s t -test). (B, C) Immunohistochemical staining of Na v 1.8 channels in the rat sciatic nerve proximal to the lesion site 48 h after ligation. The ligature was placed around the sciatic nerve proximal to the trifurcation on day 12 post-CFA. Scale bar: 100 μm. (D) Accumulation of Na v 1.8-like immunoreactivity is significantly increased in 14 day post-CFA rats compared to sham animals (* P <0.05; unpaired Student’s t- test).

Article Snippet: To detect Na v 1.8, sections were incubated with the rabbit polyclonal anti-Na v 1.8 antibody (1:200; Alomone Labs, Jerusalem, Israel) in PBS containing 0.05% Triton X-100 and 0.5% NGS.

Techniques: Expressing, Quantitative RT-PCR, Injection, Immunohistochemical staining, Staining, Ligation

Journal: Journal of Neuroinflammation

Article Title: Functional up-regulation of Na v 1.8 sodium channel in Aβ afferent fibers subjected to chronic peripheral inflammation

doi: 10.1186/1742-2094-11-45

Figure Lengend Snippet: Na v 1.8 currents are enhanced in large-sized DRG neurons from inflamed rats. (A) Immunofluorescence staining of Na v 1.8 (green) and NF200 (red) on acutely dissociated primary afferent neurons, 14 days post-CFA. Merge images show dually labeled large-sized sensory neurons (yellow). (B) Isolation of TTX-resistant Na v 1.8 currents in large-sized sensory neurons from sham and inflamed rats. Na v 1.8 currents are significantly increased post-CFA. Representative I-V curves of currents are determined using the pulse protocol indicated in the inset. (C) I-V curves of Na v 1.8 currents obtained from large-soma DRG neurons. The peak maximum current is observed at 0 mV in all groups, with the exception of day 14 (–10 mV). (D) Peak Na v 1.8 current densities are significantly increased at days 3, 8, and 14 post-CFA injection (*** P <0.001 * P <0.05 vs. sham; $$$ P <0.001 vs. day 3; ### P <0.001 vs. day 8; ANOVA followed by Sidak’s MCT; n = 6–13). (E, F) Kinetic properties of Na v 1.8 currents in large-sized sensory neurons. CFA treatment induces a leftward shift of the activation (E) and inactivation (F) curves of Na v 1.8 current. Half-activation and half-inactivation potentials and slope factors are summarized in Additional file : Table S3.

Article Snippet: To detect Na v 1.8, sections were incubated with the rabbit polyclonal anti-Na v 1.8 antibody (1:200; Alomone Labs, Jerusalem, Israel) in PBS containing 0.05% Triton X-100 and 0.5% NGS.

Techniques: Immunofluorescence, Staining, Labeling, Isolation, Injection, Activation Assay

Journal: Journal of Neuroinflammation

Article Title: Functional up-regulation of Na v 1.8 sodium channel in Aβ afferent fibers subjected to chronic peripheral inflammation

doi: 10.1186/1742-2094-11-45

Figure Lengend Snippet: Ambroxol treatment blocks the changes in the biophysical properties of Na v 1.8 in large-sized DRG neurons extracted from rats 14 days post-CFA. (A) I-V curves of Na v 1.8 currents obtained from sham and inflamed large sensory neurons. (B) Histogram showing the effects of different concentrations of ambroxol (20 and 100 μM) on the increased Na v 1.8 peak current induced by CFA (*** P <0.001, inflamed vs. sham large sensory neurons; ### P <0.001 compared to inflamed DRG neurons; ANOVA followed by Sidak’s MCT; n = 6–13). (C) Ambroxol also significantly inhibits the leftward shift of the activation curves of Na v 1.8 current. Half-activation potential and slope factor are summarized in Additional file : Table S3. Note that the data corresponding to sham and day 14 after CFA treatment are the same as the ones in Figure .

Article Snippet: To detect Na v 1.8, sections were incubated with the rabbit polyclonal anti-Na v 1.8 antibody (1:200; Alomone Labs, Jerusalem, Israel) in PBS containing 0.05% Triton X-100 and 0.5% NGS.

Techniques: Activation Assay

Journal: Frontiers in Pharmacology

Article Title: Cilostazol Ameliorates Peripheral Neuropathic Pain in Streptozotocin-Induced Type I Diabetic Rats

doi: 10.3389/fphar.2021.771271

Figure Lengend Snippet: Dose-dependent recovery of voltage-gated sodium (NaV) channel expression in dorsal root ganglia (DRG) of cilostazol-treated diabetic (DM) rats. Expressions of NaV-1.1, -1.2, -1.3, -1.6, -1.7, and -1.8 in DRG of naïve control, DM, and oral cilostazol (10, 30, and 100 mg/kg) treated DM rats were examined with (A) Western blots and (B) immunofluorescence assays. The western and immunofluorescence data revealed significant upregulation of NaV-1.1–1.7 expressions, and significant downregulation of NaV-1.8 in the DRGs of DM rats. Daily oral cilostazol treatments of 100 mg/kg, but not the lower dosages, for 6 weeks significantly reversed the DM-induced NaV dysregulation in the DRGs. Results are expressed as mean SEM for a minimum of five rats for each group. Statistical significances between the DM control and cilostazol DM treatment groups were calculated by one-way ANOVA analysis followed by the least significant difference test for multiple post hoc analyses. ** p < 0.01, * p < 0.05, *** p < 0.001.

Article Snippet: The filters were blocked with 5% milk in phosphate-buffered saline (PBS) with 0.1% Tween 20 for 1 h at room temperature and incubated for 24 h at 4 C with rabbit anti-rat Navs primary antibodies (Alomone Labs, Jerusalem, Israel) included Nav-1.1 (ASC-001), Nav-1.2 (ASC-002), Nav-1.3 (ASC-004), Nav-1.6 (ASC-009), Nav-1.7 (ASC-008), Nav-1.8 (ASC-016), and mouse anti-rat ß -actin (MilliporeSigma, MAB1501).

Techniques: Expressing, Western Blot, Immunofluorescence