charybdotoxin chtx  (Alomone Labs)

Journal: International Journal of Molecular Sciences

Article Title: The Effectiveness in Activating M-Type K+ Current Produced by Solifenacin ([(3R)-1-azabicyclo[2.2.2]octan-3-yl] (1S)-1-phenyl-3,4-dihydro-1H-isoquinoline-2-carboxylate): Independent of Its Antimuscarinic Action

doi: 10.3390/ijms222212399

Figure Lengend Snippet: Effect of SOL, SOL plus acetylcholine (ACh), SOL plus iberiotoxin (Iber), SOL plus apamin (Apa), SOL plus tolbutamide (TLB), SOL plus chlorotoxin (ChTx), SOL plus Lino, or SOL plus TRH on the amplitude of I K(M) . In these experiments, we bathed GH 3 cells in high-K + , Ca 2+ -free solution and the recording electrode was filled with K + -enriched (145 mM) solution. Current amplitude was measured at the end of the depolarizing step from −50 to −10 mV. Each bar represents the mean ± SEM ( n = 7). * Significantly different from controls ( p

Article Snippet: Chlorotoxin (ChTx) was kindly provided by Professor Dr. Woei-Jer Chuang (Department of Biochemistry, National Cheng Kung University Medical College, Tainan, Taiwan).

Techniques:

Journal: PLoS ONE

Article Title: Blockade of Kv1.3 Potassium Channels Inhibits Differentiation and Granzyme B Secretion of Human CD8+ T Effector Memory Lymphocytes

doi: 10.1371/journal.pone.0054267

Figure Lengend Snippet: K+ channel blockers inhibit GrB production by activated CD8+ T cells. Freshly isolated CD8+ T cells were pretreated with a Kv1.3 channel blocker, ShK at various concentrations (A) or with ShK (10 nM), MgTX (30 nM), ChTX (50 nM) and TRAM-34 (500 nM) (B) for 3 h, followed by stimulation with anti-CD3/CD28 or anti-CD3 alone. The levels of GrB were measured in cell supernatants by ELISA at 6 h (A) and indicated times (B and C). Data are mean of triplicate ± SD of one representative of three independent and reproducible experiments. Values that are significantly different from that of non-blocker vehicle treated control are indicated as follows: *, p

Article Snippet: Charybdotoxin (ChTx) and margatoxin (MgTx) were purchased from Alomone Labs (Jerusalem, Israel).

Techniques: Isolation, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Blockade of Kv1.3 Potassium Channels Inhibits Differentiation and Granzyme B Secretion of Human CD8+ T Effector Memory Lymphocytes

doi: 10.1371/journal.pone.0054267

Figure Lengend Snippet: Kv1.3 blockade suppresses proliferation and differentiation of anti-CD3 stimulated CD8+ T cells. ( A) Freshly isolated CD8+ T cells were pretreated with Kv channel blockers, ShK (10 nM), MgTx (30 nM) and ChTx (50 nM), for 3 hours, then stimulated with anti-CD3 alone or anti-CD3/CD28. After 4 days of culture, proliferation was measured by [3H] thymidine uptake. Data show the mean ± SD of three experiments. Significant differences are marked as follows: (*, p

Article Snippet: Charybdotoxin (ChTx) and margatoxin (MgTx) were purchased from Alomone Labs (Jerusalem, Israel).

Techniques: Isolation

Journal: PLoS ONE

Article Title: Blockade of Kv1.3 Potassium Channels Inhibits Differentiation and Granzyme B Secretion of Human CD8+ T Effector Memory Lymphocytes

doi: 10.1371/journal.pone.0054267

Figure Lengend Snippet: K+ channel blockers do not affect CD107a expression on activated CD8+ T cells. (A) Freshly isolated CD8+ T cells were stimulated with anti-CD3/CD28 or anti-CD3 for 24 hours. Cells were then stained with a CD107a-specific mAb, or an IgG1 isotype control (filled histogram) at the indicated times. (B). CD8+ T cells were pretreated with ShK (10 nM), MgTX (30 nM), ChTX (50 nM) and TRAM-34 (500 nM) for 3 h, followed by stimulation with anti-CD3/CD28 or anti-CD3 alone for 6 hours. Surface expression levels of CD107a were then analyzed by flow cytometry. FACS plots shown are representative data from three separate experiments.

Article Snippet: Charybdotoxin (ChTx) and margatoxin (MgTx) were purchased from Alomone Labs (Jerusalem, Israel).

Techniques: Expressing, Isolation, Staining, Flow Cytometry, Cytometry, FACS

Journal: PLoS ONE

Article Title: Physiological Epidermal Growth Factor Concentrations Activate High Affinity Receptors to Elicit Calcium Oscillations

doi: 10.1371/journal.pone.0106803

Figure Lengend Snippet: External Ca 2+ dependence and sensitivity to K + channel blocker charybdotoxin of EGF Ca 2+ transients. A/Proportion of cells responding (grey bar) or not responding (white bar) to 2 nM EGF in 3 mM extracellular Ca 2+ (n = 24) or in 0 mM Ca 2+/ 1 mM EGTA (n = 28) in the extracellular medium. B/Fluorescence intensity signaling of individual cells (each represented by a different color) during the application of 2 nM EGF (white bar) when 3 mM Ca 2+ was present (n = 24). The averaged population signal is shown as a thick black trace. C/Fluorescence intensity of individual cells (each represented by a different color) during the application of 2 nM EGF (white bar) when Ca 2+ was removed and 1 mM EGTA was added to the extracellular medium (n = 28). The averaged population signal is shown as a thick black trace. D/Average of all cell signals during 2 nM EGF application, synchronized at the time of the first fluorescence peak and averaged for 150 sec, when 3 mM Ca 2+ was present (black line, n = 24) or when Ca 2+ was removed from and 1 mM EGTA was added to the extracellular medium (red line, n = 28). E/Proportion of cells responding (grey bar) or not responding (white bar) to 20 pM EGF in 3 mM extracellular Ca 2+ (n = 13) or in 0 mM Ca 2+/ 1 mM EGTA (n = 11) in the extracellular medium. F/Fluorescence intensity of individual cells (each represented by a different color) during the application of 20 pM EGF (white bar) when 3 mM Ca 2+ was present (n = 13). The averaged population signal is shown as a thick black trace. G/Fluorescence intensity of individual cells (each represented by a different color) during the application of 20 pM EGF (white bar) when Ca 2+ was removed from and 1 mM EGTA was added to the extracellular medium (n = 11). The averaged population signal is shown as a thick black trace. H/Average of all cell signals during 20 pM EGF application, synchronized at the time the first fluorescence peak and for 150 sec, when 3 mM Ca 2+ was present (black line, n = 13) or when Ca 2+ was removed from and 1 mM EGTA was added to the extracellular medium (red line, n = 11). I/Proportion of cells responding (grey bar) or not responding (white bar) to 2 nM EGF in the absence (0, n = 24/27) or in the presence (100, n = 16/19) of 100 nM charybdotoxin (chx) in the extracellular medium. J/Proportion of cells responding (grey bar) or not responding (white bar) to 20 pM EGF in the absence (0, n = 16/22) or in the presence (100, n = 6/22) of 100 nM charybdotoxin (chx) in the extracellular medium.

Article Snippet: Charybdotoxin (Alomone labs), a blocker of Ca2+ -activated K+ channels KCa1.1 , KCa3.1 and voltage-dependent Kv1.3 channels was added to cells at a concentration of 100 nM, 20 min before starting the time-lapse recording.

Techniques: Fluorescence, Size-exclusion Chromatography

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: HIV-1 gp120 and chemokines activate ion channels in primary macrophages through CCR5 and CXCR4 stimulation

doi:

Figure Lengend Snippet: Potassium and chloride currents evoked by gp120 in MDM. The ionic nature of currents elicited in MDM by gp120 from JRFL ( A ) and IIIB ( B ) was determined from the reversal potential (right) and with channel inhibitors (left). At peak activation, the outward JRFL current reversed direction at ≈−78 mV ( A , top right) whereas the peak inward currents elicited by JRFL ( A , bottom right) and IIIB ( B , right) reversed at ≈5 mV (squares). In low Cl − bath solution, the current voltage relationship of the inward current elicited by both Envs shifted to ≈+40 mV (circles). Charybdotoxin (100 nM) blocked the JRFL-evoked outward current, and NPPB (10 μM) blocked the inward currents activated by both Envs ( A and B , left).

Article Snippet: The pharmacological inhibitors NPPB, IAA, niflumic acid, and DIDS were obtained from Calbiochem, and charybdotoxin was obtained from Alomone Laboratories (Jerusalem).

Techniques: Activation Assay