anti kcnq2  (Alomone Labs)

Journal: International Journal of Molecular Sciences

Article Title: Proteomic-Based Approach Reveals the Involvement of Apolipoprotein A-I in Related Phenotypes of Autism Spectrum Disorder in the BTBR Mouse Model

doi: 10.3390/ijms232315290

Figure Lengend Snippet: ApoA-I-related pathway regulates the expressions of proteins related to the apoptosis process and the KCNQ2 channel in the hippocampus of the mice. ( a , b ) Representative Western blotting bands and quantification of the apoptosis proteins, CDK5/β-tubulin, caspase-3/β-tubulin, Bax/β-tubulin and Bcl-2/GAPDH ratios. ( c ) Quantification of mRNA expression of KCND2, KCND3 and KCNJ10 channels. ( d ) Quantification of KCNQ2, and KCNQ3 mRNA expression. ( e , f ) Representative Western blotting bands and quantification of the M channel proteins, KCNQ2/GAPDH, and KCNQ3/GAPDH ratios. N = 8–10 per group. All data are shown in bar diagrams, which reflect the arithmetic mean ± standard error of the mean. * p < 0.05, ** p < 0.01.

Article Snippet: The antibodies used in this experiment were listed as follows: ApoA-I (Cat. 66206-1-AP, RRID: AB_2881597, 1:2000, Proteintech, Rosemont, IL, USA), SphK1 (Cat. 10670-1-AP, RRID: AB_2195809, 1:1000, Proteintech), SphK2 (Cat. 17096-1-AP, RRID: AB_10598479, 1:3000, Proteintech), phospho-p38 MAPK (Cat. 4511S, RRID: AB_2139682, 1:1000; Cell Signaling, Danvers, MA, USA), p38 MAPK (Cat. 8690S, RRID: AB_10999090, 1:1000, Cell Signaling), phospho-p44/42 MAPK (Cat. 4370S, RRID: AB_2315112, 1:2000, Cell Signaling), p44/42 MAPK (Erk1/2) (Cat. 4695s, RRID: AB_390779, 1:2000, Cell Signaling), CDK5 (D1F7M) (Cat. 14145s, RRID: AB_2773717, 1:1000, Cell Signaling), GAD1 (D1F2M) (Cat. 41318s, RRID: AB_2799198, 1:1000, Cell Signaling), Bax (D3R2M) (Cat. 14796s, RRID: AB_2716251, 1:1000, Cell Signaling), Bcl-2 (D17C4) (Cat. 3498s, RRID: AB_1903907, 1:1000; Cell Signaling), caspase-3 (Cat. 14220s, RRID: AB_2798429, 1:1000, Cell Signaling), phospho-CaMKII (Thr286) (D21E4) (Cat. 12716s, RRID: AB_2713889, 1:1000, Cell Signaling), CaMKII-α (D10C11) (Cat. 11945s, RRID: AB_2797775, 1:1000, Cell Signaling), CREB (Cat. 9197s, RRID: AB_331277, 1:1000, Cell Signaling), phospho-CREB (Cat. 9198s, RRID: AB_2561044, 1:1000, Cell Signaling), KCNQ2 (Cat. AGP-065, RRID: AB_2827304, 1:200; Alomone), KCNQ3 (Cat. APC-051, RRID: AB_2040103, 1:200; Alomone), KCNQ5 (Cat. APC-155, RRID: AB_2341038, 1:200; Alomone) β-actin (Cat. AC026, RRID: AB_2768234, 1:50000, ABclonal), β-tubulin (Cat. 10094-1 AP, RRID: AB_2210695, 1:5000, Proteintech) and GAPDH (Cat. 10494-1-AP, RRID: AB_2263076, 1:5000, Proteintech).

Techniques: Western Blot, Expressing

Journal: Science (New York, N.Y.)

Article Title: Hyperexcitable arousal circuits drive sleep instability during aging

doi: 10.1126/science.abh3021

Figure Lengend Snippet: (A) Representative traces from a young Hcrt neuron (left) before or (right) in the presence of a KCNQ2/3 blocker XE991 (50 μM). (B) XE991 depolarized RMP and (C) increased firing frequency in young Hcrt neurons (n = 19 neurons from seven mice). (D) Representative traces from an aged Hcrt neuron (left) before or (right) in the presence of a KCNQ2/3 activator flupirtine (50 μM). (E) Flupirtine hyperpolarized RMP and (F) decreased firing frequency in aged Hcrt neurons (n = 8 neurons from five mice). (G) IM in young Hcrt neurons modulated by XE991 (top; n = 6 neurons from three mice) and flupirtine (bottom; n = 10 neurons from five mice). (H) IM in aged Hcrt neurons modulated by XE991 (top; n = 7 neurons from three mice) and flupirtine (bottom; n = 15 neurons from five mice). (I) Basal IM in young Hcrt neurons (n = 25 neurons from nine mice) versus in aged Hcrt neurons (n = 26 neurons from nine mice). (J) Array tomography revealed reduced KCNQ2 expression in aged Hcrt neurons (n = 4 mice/group). In (B) and (C), Wilcoxon matched-pairs signed rank test; (E) and (F), RM one-way ANOVA followed by post hoc Tukey’s multiple comparisons; (G), (H), and (J), paired t test; (I) unpaired t test with Welch’s correction; statistical details are available in the supplementary text.

Article Snippet: Immunohistochemistry Immunohistochemistry was then carried out on the arrays using primary antibody against KCNQ2 (Alomone Cat. no. AGP-065).

Techniques: Tomography, Expressing

Journal: Science (New York, N.Y.)

Article Title: Hyperexcitable arousal circuits drive sleep instability during aging

doi: 10.1126/science.abh3021

Figure Lengend Snippet: (A) Significantly increased wake amount by the KCNQ2/3 blocker XE991 (2 mg/kg) in young mice. (B) Significantly increased NREM amount and mean bout length by the KCNQ2/3 activator flupirtine (20 mg/kg) in aged mice. (C) Representative EMG-EEG raw traces from vehicle- and XE991-treated (2 mg/kg) young mice. (D) Representative EMG-EEG raw traces from vehicle- and flupirtine-treated (20 mg/kg) aged mice. (E) Power spectra of EEG for vehicle- and XE991-treated young mice and (F) power spectra of EEG for vehicle- and flupirtine-treated aged mice. (G) Comparison of delta, theta band power between vehicle- and XE991-treated young mice and (H) between vehicle- and flupirtine-treated aged mice. Data indicate mean ± SEM [young, n = 7 mice each group; aged, n = 6 mice each group; (A) and (B) two-way liner mixed-effects model followed by Šidák’s multiple comparisons, dark phase indicated by gray shielding; (G) and (H) Holm-Šidák, *P < 0.05, ***P < 0.005, ****P < 0.001, †P < 0.0005; statistical details are available in the supplementary text].

Article Snippet: Immunohistochemistry Immunohistochemistry was then carried out on the arrays using primary antibody against KCNQ2 (Alomone Cat. no. AGP-065).

Techniques: