fitc  (Alomone Labs)


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    Structured Review

    Alomone Labs fitc
    Fitc, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fitc/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    fitc - by Bioz Stars, 2022-10
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    Alomone Labs anti kv1 5
    Characterization of the Kv1.3 and <t>Kv1.5</t> mutant channels containing the YS segment. A , average normalized activation and inactivation curves are shown as conductance-voltage relationships for Kv1.3, Kv1.5, the truncated Kv1.3-YS channel, and the chimeras Kv1.5-YS 532 and Kv1.5-YS 613 . All datasets were fitted to Boltzmann functions. Each data point is the mean ± S.E. of 6–11 cells. B , confocal images of non-permeabilized cells transfected with Kv1.3-YS-Cherry, Kv1.5-YS 532 -EGFP, and Kv1.5-YS 613 -EGFP. An extracellular anti-Kv1.3 antibody was used to label Kv1.3-YS ( green ), whereas the extracellular anti-Kv1.5 antibody was used for Kv1.5-YS 532 and Kv1.5-YS 613 chimeras ( red ). Nuclei were stained by Hoechst ( blue ). C , proliferation rate of the indicated channels or GFP-transfected cells (control) was determined by measuring EdU incorporation. Significant differences when comparing to Kv1.3 (*) or to control (#) are indicated. Statistical analysis was performed with one-way ANOVA followed by a Tukey's HSD multiple comparison. Each bar is the average of 9–15 determinations from 5 different assays. D , the average peak current amplitude obtained in cell-attached experiments for Kv1.5 channels and all the Kv1.5 chimeras was plotted against the % of the channels expressed at the plasma membrane ( upper graph ) or their normalized effect on proliferation (taking 100% as the proliferation rate of GFP-transfected HEK cells, lower graph ). The correlation between expression and current was fit to a linear regression curve ( y = 18.54 + 0.0066x, R 2 = 0.85, p = 0.008), but there was no correlation between proliferation and current amplitude ( R 2 = 0.23, p = 0.19).
    Anti Kv1 5, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti kv1 5/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    93
    Alomone Labs anti p2y6 receptor fitc
    Decrease of <t>P2Y6</t> Receptor and Calcium mobilization after treatment without change in intracellular PAD4 levels. (A) Neutrophils were treated as described before. P2Y6 was measured by flow cytometry with a <t>FITC-conjugated</t> antibody. Treatment with Alveofact ® decreases extracellular expression of the P2Y6 receptor independent of timing or dosing, while Curosurf ® does not exhibit this effect in every dosing. (B) Calcium levels at 3 h after treatment were used. Data were normalized to stimulated PMA control due to the heterogeneity in basic calcium levels and the response of the neutrophils to stimulation. Results are similar to the findings of the P2Y6 receptor expression. (C) Intracellular PAD4 levels were determined by ELISA using cell lysates of 1×10 6 cells. Interestingly, PAD4 levels did not change after treatment with surfactant compared to PMA-stimulated neutrophils. Significance level was set as p
    Anti P2y6 Receptor Fitc, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p2y6 receptor fitc/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    Image Search Results


    Characterization of the Kv1.3 and Kv1.5 mutant channels containing the YS segment. A , average normalized activation and inactivation curves are shown as conductance-voltage relationships for Kv1.3, Kv1.5, the truncated Kv1.3-YS channel, and the chimeras Kv1.5-YS 532 and Kv1.5-YS 613 . All datasets were fitted to Boltzmann functions. Each data point is the mean ± S.E. of 6–11 cells. B , confocal images of non-permeabilized cells transfected with Kv1.3-YS-Cherry, Kv1.5-YS 532 -EGFP, and Kv1.5-YS 613 -EGFP. An extracellular anti-Kv1.3 antibody was used to label Kv1.3-YS ( green ), whereas the extracellular anti-Kv1.5 antibody was used for Kv1.5-YS 532 and Kv1.5-YS 613 chimeras ( red ). Nuclei were stained by Hoechst ( blue ). C , proliferation rate of the indicated channels or GFP-transfected cells (control) was determined by measuring EdU incorporation. Significant differences when comparing to Kv1.3 (*) or to control (#) are indicated. Statistical analysis was performed with one-way ANOVA followed by a Tukey's HSD multiple comparison. Each bar is the average of 9–15 determinations from 5 different assays. D , the average peak current amplitude obtained in cell-attached experiments for Kv1.5 channels and all the Kv1.5 chimeras was plotted against the % of the channels expressed at the plasma membrane ( upper graph ) or their normalized effect on proliferation (taking 100% as the proliferation rate of GFP-transfected HEK cells, lower graph ). The correlation between expression and current was fit to a linear regression curve ( y = 18.54 + 0.0066x, R 2 = 0.85, p = 0.008), but there was no correlation between proliferation and current amplitude ( R 2 = 0.23, p = 0.19).

    Journal: The Journal of Biological Chemistry

    Article Title: Molecular Determinants of Kv1.3 Potassium Channels-induced Proliferation *

    doi: 10.1074/jbc.M115.678995

    Figure Lengend Snippet: Characterization of the Kv1.3 and Kv1.5 mutant channels containing the YS segment. A , average normalized activation and inactivation curves are shown as conductance-voltage relationships for Kv1.3, Kv1.5, the truncated Kv1.3-YS channel, and the chimeras Kv1.5-YS 532 and Kv1.5-YS 613 . All datasets were fitted to Boltzmann functions. Each data point is the mean ± S.E. of 6–11 cells. B , confocal images of non-permeabilized cells transfected with Kv1.3-YS-Cherry, Kv1.5-YS 532 -EGFP, and Kv1.5-YS 613 -EGFP. An extracellular anti-Kv1.3 antibody was used to label Kv1.3-YS ( green ), whereas the extracellular anti-Kv1.5 antibody was used for Kv1.5-YS 532 and Kv1.5-YS 613 chimeras ( red ). Nuclei were stained by Hoechst ( blue ). C , proliferation rate of the indicated channels or GFP-transfected cells (control) was determined by measuring EdU incorporation. Significant differences when comparing to Kv1.3 (*) or to control (#) are indicated. Statistical analysis was performed with one-way ANOVA followed by a Tukey's HSD multiple comparison. Each bar is the average of 9–15 determinations from 5 different assays. D , the average peak current amplitude obtained in cell-attached experiments for Kv1.5 channels and all the Kv1.5 chimeras was plotted against the % of the channels expressed at the plasma membrane ( upper graph ) or their normalized effect on proliferation (taking 100% as the proliferation rate of GFP-transfected HEK cells, lower graph ). The correlation between expression and current was fit to a linear regression curve ( y = 18.54 + 0.0066x, R 2 = 0.85, p = 0.008), but there was no correlation between proliferation and current amplitude ( R 2 = 0.23, p = 0.19).

    Article Snippet: Non-permeabilized cells were incubated with anti-Kv1.3 or anti Kv1.5 extracellular primary antibodies (APC101 or APC150, Alomone Labs), whereas permeabilized cells were incubated with anti-Kv1.3 COOH (75-009, NeuroMab) or anti-Kv1.5 COOH (APC004, Alomone Labs), all at a final concentration of 1:50.

    Techniques: Mutagenesis, Activation Assay, Transfection, Staining, Expressing

    Mutant KCNA5 is located in perinuclear packets and not on the cell surface of transfected HEK-293 cells and human (h)PASMC. A : HEK-293 cells transfected with WT KCNA5 ( a and c ) or G182R ( b and d ) were stained with anti-KCNA5 antibody (Ab-KCNA5, red) and

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Tetramerization domain mutations in KCNA5 affect channel kinetics and cause abnormal trafficking patterns

    doi: 10.1152/ajpcell.00464.2009

    Figure Lengend Snippet: Mutant KCNA5 is located in perinuclear packets and not on the cell surface of transfected HEK-293 cells and human (h)PASMC. A : HEK-293 cells transfected with WT KCNA5 ( a and c ) or G182R ( b and d ) were stained with anti-KCNA5 antibody (Ab-KCNA5, red) and

    Article Snippet: After cells were fixed in 4% paraformaldehyde-PBS for 15 min at room temperature, coverslips were incubated in blocking solution (2% BSA, 2% FBS, 0.1% Triton X-100 in PBS) for 1 h. Coverslips were then exposed to rabbit anti-KV1.5 antibody (Alomone Labs) at a dilution of 1:100 in blocking solution for 2 h at room temperature.

    Techniques: Mutagenesis, Transfection, Staining

    G182R and E211D mutations cause incomplete processing of KCNA5 in HEK-293 cells. HEK-293 cells were transfected with WT KCNA5, G182R, E211D, or G182R/E211D and subjected to standard immunoblot procedures. A : representative immunoblot from cells transfected

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Tetramerization domain mutations in KCNA5 affect channel kinetics and cause abnormal trafficking patterns

    doi: 10.1152/ajpcell.00464.2009

    Figure Lengend Snippet: G182R and E211D mutations cause incomplete processing of KCNA5 in HEK-293 cells. HEK-293 cells were transfected with WT KCNA5, G182R, E211D, or G182R/E211D and subjected to standard immunoblot procedures. A : representative immunoblot from cells transfected

    Article Snippet: After cells were fixed in 4% paraformaldehyde-PBS for 15 min at room temperature, coverslips were incubated in blocking solution (2% BSA, 2% FBS, 0.1% Triton X-100 in PBS) for 1 h. Coverslips were then exposed to rabbit anti-KV1.5 antibody (Alomone Labs) at a dilution of 1:100 in blocking solution for 2 h at room temperature.

    Techniques: Transfection

    Mutant KCNA5 forms functional homotetrameric channels. A : COS-1, HEK-293, and human pulmonary artery smooth muscle cells (PASMC) were transiently transfected with either empty vector [green fluorescent protein (GFP)] or WT KCNA5 (WT) as indicated. Whole

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Tetramerization domain mutations in KCNA5 affect channel kinetics and cause abnormal trafficking patterns

    doi: 10.1152/ajpcell.00464.2009

    Figure Lengend Snippet: Mutant KCNA5 forms functional homotetrameric channels. A : COS-1, HEK-293, and human pulmonary artery smooth muscle cells (PASMC) were transiently transfected with either empty vector [green fluorescent protein (GFP)] or WT KCNA5 (WT) as indicated. Whole

    Article Snippet: After cells were fixed in 4% paraformaldehyde-PBS for 15 min at room temperature, coverslips were incubated in blocking solution (2% BSA, 2% FBS, 0.1% Triton X-100 in PBS) for 1 h. Coverslips were then exposed to rabbit anti-KV1.5 antibody (Alomone Labs) at a dilution of 1:100 in blocking solution for 2 h at room temperature.

    Techniques: Mutagenesis, Functional Assay, Transfection, Plasmid Preparation

    Mutations in KCNA5 at G182 and E211 do not affect the pharmacological effect of 4-aminopyridine (4-AP). A : a standard I-V pulse protocol was delivered to HEK-293 cells transiently transfected with the indicated vector [WT KCNA5 ( a ), G182R ( b ), E211D (

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Tetramerization domain mutations in KCNA5 affect channel kinetics and cause abnormal trafficking patterns

    doi: 10.1152/ajpcell.00464.2009

    Figure Lengend Snippet: Mutations in KCNA5 at G182 and E211 do not affect the pharmacological effect of 4-aminopyridine (4-AP). A : a standard I-V pulse protocol was delivered to HEK-293 cells transiently transfected with the indicated vector [WT KCNA5 ( a ), G182R ( b ), E211D (

    Article Snippet: After cells were fixed in 4% paraformaldehyde-PBS for 15 min at room temperature, coverslips were incubated in blocking solution (2% BSA, 2% FBS, 0.1% Triton X-100 in PBS) for 1 h. Coverslips were then exposed to rabbit anti-KV1.5 antibody (Alomone Labs) at a dilution of 1:100 in blocking solution for 2 h at room temperature.

    Techniques: Transfection, Plasmid Preparation

    Decreased G182R expression in COS-1 cells cannot be rescued by overexpression of K V β subunits. A , a : HEK-293 cells were transfected with WT KCNA5 or K V β1.3-HA alone or cotransfected with WT KCNA5, G182R, E211D, or G182R/E211D and K V β1.3-HA.

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Tetramerization domain mutations in KCNA5 affect channel kinetics and cause abnormal trafficking patterns

    doi: 10.1152/ajpcell.00464.2009

    Figure Lengend Snippet: Decreased G182R expression in COS-1 cells cannot be rescued by overexpression of K V β subunits. A , a : HEK-293 cells were transfected with WT KCNA5 or K V β1.3-HA alone or cotransfected with WT KCNA5, G182R, E211D, or G182R/E211D and K V β1.3-HA.

    Article Snippet: After cells were fixed in 4% paraformaldehyde-PBS for 15 min at room temperature, coverslips were incubated in blocking solution (2% BSA, 2% FBS, 0.1% Triton X-100 in PBS) for 1 h. Coverslips were then exposed to rabbit anti-KV1.5 antibody (Alomone Labs) at a dilution of 1:100 in blocking solution for 2 h at room temperature.

    Techniques: Expressing, Over Expression, Transfection

    Two nonsynonymous mutations identified in the KCNA5 gene from idiopathic pulmonary arterial hypertension (IPAH) patients localize to the NH 2 -terminal tetramerization domain (T1 domain). A , left : schematic diagram of voltage-gated K + (K V ) channel subunit

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Tetramerization domain mutations in KCNA5 affect channel kinetics and cause abnormal trafficking patterns

    doi: 10.1152/ajpcell.00464.2009

    Figure Lengend Snippet: Two nonsynonymous mutations identified in the KCNA5 gene from idiopathic pulmonary arterial hypertension (IPAH) patients localize to the NH 2 -terminal tetramerization domain (T1 domain). A , left : schematic diagram of voltage-gated K + (K V ) channel subunit

    Article Snippet: After cells were fixed in 4% paraformaldehyde-PBS for 15 min at room temperature, coverslips were incubated in blocking solution (2% BSA, 2% FBS, 0.1% Triton X-100 in PBS) for 1 h. Coverslips were then exposed to rabbit anti-KV1.5 antibody (Alomone Labs) at a dilution of 1:100 in blocking solution for 2 h at room temperature.

    Techniques:

    Cotransfection of K V β subunits affects KCNA5 channel kinetics. HEK-293 cells were transfected with WT KCNA5 alone (KCNA5) or in the presence of K V β1.3-hemagglutinin (HA) (KCNA5/K V β1.3). A and B : representative current recordings

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Tetramerization domain mutations in KCNA5 affect channel kinetics and cause abnormal trafficking patterns

    doi: 10.1152/ajpcell.00464.2009

    Figure Lengend Snippet: Cotransfection of K V β subunits affects KCNA5 channel kinetics. HEK-293 cells were transfected with WT KCNA5 alone (KCNA5) or in the presence of K V β1.3-hemagglutinin (HA) (KCNA5/K V β1.3). A and B : representative current recordings

    Article Snippet: After cells were fixed in 4% paraformaldehyde-PBS for 15 min at room temperature, coverslips were incubated in blocking solution (2% BSA, 2% FBS, 0.1% Triton X-100 in PBS) for 1 h. Coverslips were then exposed to rabbit anti-KV1.5 antibody (Alomone Labs) at a dilution of 1:100 in blocking solution for 2 h at room temperature.

    Techniques: Cotransfection, Transfection

    Voltage-dependent inactivation is accelerated in the G182R mutant KCNA5 channel. A standard 2-pulse inactivation protocol was used to determine channel availability after a 10-s prepulse in HEK-293 cells transiently transfected with WT KCNA5, G182R, E211D,

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Tetramerization domain mutations in KCNA5 affect channel kinetics and cause abnormal trafficking patterns

    doi: 10.1152/ajpcell.00464.2009

    Figure Lengend Snippet: Voltage-dependent inactivation is accelerated in the G182R mutant KCNA5 channel. A standard 2-pulse inactivation protocol was used to determine channel availability after a 10-s prepulse in HEK-293 cells transiently transfected with WT KCNA5, G182R, E211D,

    Article Snippet: After cells were fixed in 4% paraformaldehyde-PBS for 15 min at room temperature, coverslips were incubated in blocking solution (2% BSA, 2% FBS, 0.1% Triton X-100 in PBS) for 1 h. Coverslips were then exposed to rabbit anti-KV1.5 antibody (Alomone Labs) at a dilution of 1:100 in blocking solution for 2 h at room temperature.

    Techniques: Mutagenesis, Transfection

    G182R protein expression is significantly decreased in COS-1 cells. A : COS-1 cells were transiently transfected with water (Mock), WT-KCNA5, G182R, E211D, or G182R/E211D. Representative images are shown at ×40 magnification. B : transfected cells

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Tetramerization domain mutations in KCNA5 affect channel kinetics and cause abnormal trafficking patterns

    doi: 10.1152/ajpcell.00464.2009

    Figure Lengend Snippet: G182R protein expression is significantly decreased in COS-1 cells. A : COS-1 cells were transiently transfected with water (Mock), WT-KCNA5, G182R, E211D, or G182R/E211D. Representative images are shown at ×40 magnification. B : transfected cells

    Article Snippet: After cells were fixed in 4% paraformaldehyde-PBS for 15 min at room temperature, coverslips were incubated in blocking solution (2% BSA, 2% FBS, 0.1% Triton X-100 in PBS) for 1 h. Coverslips were then exposed to rabbit anti-KV1.5 antibody (Alomone Labs) at a dilution of 1:100 in blocking solution for 2 h at room temperature.

    Techniques: Expressing, Transfection

    The experimental steps showing the inhibitory effect of potassium channels on microglial migration. ( A ) The monolayer of BV2 microglial cells was scratched with a sterile pipette tip and incubated with the medium or the pharmacological inhibitors or the cells were transfected with small interfering RNA (siRNA); pictures were analyzed at t24h and the inhibition of Kv1.3 and Kir2.1 reduced the rate of migration in the BV2 microglial cell line. ( B ) CX3CR1-eGFP transgenic mice were subjected to SNI or sham surgeries, the ipsilateral spinal cord dorsal horn (SC-DH) was dissected and cultured for 3 h in 8 µm pore inserts, in the culture medium or in the presence of the inhibitors, and the rate of migration was quantified in each condition. Different mechanisms by which the inhibition of the investigated potassium channels may influence the microglial migration rate, are proposed in the outlined box. PBS: phosphate buffer saline; PFA: paraformaldehyde.

    Journal: International Journal of Molecular Sciences

    Article Title: Potassium Channels Kv1.3 and Kir2.1 But Not Kv1.5 Contribute to BV2 Cell Line and Primary Microglial Migration

    doi: 10.3390/ijms22042081

    Figure Lengend Snippet: The experimental steps showing the inhibitory effect of potassium channels on microglial migration. ( A ) The monolayer of BV2 microglial cells was scratched with a sterile pipette tip and incubated with the medium or the pharmacological inhibitors or the cells were transfected with small interfering RNA (siRNA); pictures were analyzed at t24h and the inhibition of Kv1.3 and Kir2.1 reduced the rate of migration in the BV2 microglial cell line. ( B ) CX3CR1-eGFP transgenic mice were subjected to SNI or sham surgeries, the ipsilateral spinal cord dorsal horn (SC-DH) was dissected and cultured for 3 h in 8 µm pore inserts, in the culture medium or in the presence of the inhibitors, and the rate of migration was quantified in each condition. Different mechanisms by which the inhibition of the investigated potassium channels may influence the microglial migration rate, are proposed in the outlined box. PBS: phosphate buffer saline; PFA: paraformaldehyde.

    Article Snippet: The potassium channels were visualized with primary antibodies anti-Kv1.3, anti-Kv1.5, anti-Kir2.1 (#APC-002, #APC-004, #APC-0026 Alomone, Jerusalem, Israel), used in a dilution of 1:300 and the secondary antibody was goat anti-rabbit Alexa Fluor 568 (1:1500, #A11011, Life Technologies, Carlsbad, CA, USA).

    Techniques: Migration, Transferring, Incubation, Transfection, Small Interfering RNA, Inhibition, Transgenic Assay, Mouse Assay, Cell Culture

    Immunolabeling of potassium channels in cultured BV2 microglial cells. The cells were stained with antibodies targeting Kv1.3 ( A ), Kv1.5 ( B ), Kir2.1 ( C ) and the negative control with Alexa 568 ( D ). The coverslips were mounted using the Prolong antifade with DAPI in blue. The channel was pseudo-colored in green for improved visualization. The images are representative of three independent experiments. Scale bar: 40 µm.

    Journal: International Journal of Molecular Sciences

    Article Title: Potassium Channels Kv1.3 and Kir2.1 But Not Kv1.5 Contribute to BV2 Cell Line and Primary Microglial Migration

    doi: 10.3390/ijms22042081

    Figure Lengend Snippet: Immunolabeling of potassium channels in cultured BV2 microglial cells. The cells were stained with antibodies targeting Kv1.3 ( A ), Kv1.5 ( B ), Kir2.1 ( C ) and the negative control with Alexa 568 ( D ). The coverslips were mounted using the Prolong antifade with DAPI in blue. The channel was pseudo-colored in green for improved visualization. The images are representative of three independent experiments. Scale bar: 40 µm.

    Article Snippet: The potassium channels were visualized with primary antibodies anti-Kv1.3, anti-Kv1.5, anti-Kir2.1 (#APC-002, #APC-004, #APC-0026 Alomone, Jerusalem, Israel), used in a dilution of 1:300 and the secondary antibody was goat anti-rabbit Alexa Fluor 568 (1:1500, #A11011, Life Technologies, Carlsbad, CA, USA).

    Techniques: Immunolabeling, Cell Culture, Staining, Negative Control

    The contribution of potassium channels in BV2 microglial migration. The representative images show the robust and repetitive scratch made with the 200 µL sterile pipette tip at t0h ( A ) and the cell migration in control conditions ( E ) and after the inhibitor at t24h ( F – H ). The histograms show that BV2 microglial cells migrate less after the inhibition of Kv1.3 and Kir2.1 ( B , D ), whereas blocking Kv1.5 has no effect on cellular migration ( C ). Scale bar: 100 µm. All the statistical analysis is represented as Mean ± SD, ***: p

    Journal: International Journal of Molecular Sciences

    Article Title: Potassium Channels Kv1.3 and Kir2.1 But Not Kv1.5 Contribute to BV2 Cell Line and Primary Microglial Migration

    doi: 10.3390/ijms22042081

    Figure Lengend Snippet: The contribution of potassium channels in BV2 microglial migration. The representative images show the robust and repetitive scratch made with the 200 µL sterile pipette tip at t0h ( A ) and the cell migration in control conditions ( E ) and after the inhibitor at t24h ( F – H ). The histograms show that BV2 microglial cells migrate less after the inhibition of Kv1.3 and Kir2.1 ( B , D ), whereas blocking Kv1.5 has no effect on cellular migration ( C ). Scale bar: 100 µm. All the statistical analysis is represented as Mean ± SD, ***: p

    Article Snippet: The potassium channels were visualized with primary antibodies anti-Kv1.3, anti-Kv1.5, anti-Kir2.1 (#APC-002, #APC-004, #APC-0026 Alomone, Jerusalem, Israel), used in a dilution of 1:300 and the secondary antibody was goat anti-rabbit Alexa Fluor 568 (1:1500, #A11011, Life Technologies, Carlsbad, CA, USA).

    Techniques: Migration, Transferring, Inhibition, Blocking Assay

    Electrophysiological properties of BV2 cells. For all Kv1.3, Kv1.5 and Kir2.1 potassium channels, the electrophysiological characteristics are represented by the intensity/voltage (I/V) curves extracted from the step recordings, representing the total current elicited by a voltage protocol starting from −120 mV, in 10 mV increments, to +40 mV ( A – C ), in control conditions and after each specific inhibitor. The inhibition effect of the of each blocker can be seen from the I/V curves. The insets represent the comparison of the current amplitude elicited by a voltage step at −160 mV and +40 mV. All the data are represented as Mean ± SD ( n = 3 independent experiments).

    Journal: International Journal of Molecular Sciences

    Article Title: Potassium Channels Kv1.3 and Kir2.1 But Not Kv1.5 Contribute to BV2 Cell Line and Primary Microglial Migration

    doi: 10.3390/ijms22042081

    Figure Lengend Snippet: Electrophysiological properties of BV2 cells. For all Kv1.3, Kv1.5 and Kir2.1 potassium channels, the electrophysiological characteristics are represented by the intensity/voltage (I/V) curves extracted from the step recordings, representing the total current elicited by a voltage protocol starting from −120 mV, in 10 mV increments, to +40 mV ( A – C ), in control conditions and after each specific inhibitor. The inhibition effect of the of each blocker can be seen from the I/V curves. The insets represent the comparison of the current amplitude elicited by a voltage step at −160 mV and +40 mV. All the data are represented as Mean ± SD ( n = 3 independent experiments).

    Article Snippet: The potassium channels were visualized with primary antibodies anti-Kv1.3, anti-Kv1.5, anti-Kir2.1 (#APC-002, #APC-004, #APC-0026 Alomone, Jerusalem, Israel), used in a dilution of 1:300 and the secondary antibody was goat anti-rabbit Alexa Fluor 568 (1:1500, #A11011, Life Technologies, Carlsbad, CA, USA).

    Techniques: Inhibition

    BV2 microglial migration after the silencing of the Kv1.3 and Kir2.1 channels. The representative images show the scratch made with the 200 µL sterile pipette tip at t0h ( A ) and the cell migration after scrambled and silencing siRNA at t24h ( B – D ) and at t48h ( E – G ). The bar graphs show the BV2 migration after scrambled and silencing siRNA at t24h ( H ) and at t48h ( I ). Scale bar: 100 µm. All the statistical analysis are represented as Mean ± SD, ***: p

    Journal: International Journal of Molecular Sciences

    Article Title: Potassium Channels Kv1.3 and Kir2.1 But Not Kv1.5 Contribute to BV2 Cell Line and Primary Microglial Migration

    doi: 10.3390/ijms22042081

    Figure Lengend Snippet: BV2 microglial migration after the silencing of the Kv1.3 and Kir2.1 channels. The representative images show the scratch made with the 200 µL sterile pipette tip at t0h ( A ) and the cell migration after scrambled and silencing siRNA at t24h ( B – D ) and at t48h ( E – G ). The bar graphs show the BV2 migration after scrambled and silencing siRNA at t24h ( H ) and at t48h ( I ). Scale bar: 100 µm. All the statistical analysis are represented as Mean ± SD, ***: p

    Article Snippet: The potassium channels were visualized with primary antibodies anti-Kv1.3, anti-Kv1.5, anti-Kir2.1 (#APC-002, #APC-004, #APC-0026 Alomone, Jerusalem, Israel), used in a dilution of 1:300 and the secondary antibody was goat anti-rabbit Alexa Fluor 568 (1:1500, #A11011, Life Technologies, Carlsbad, CA, USA).

    Techniques: Migration, Transferring

    The migration of primary microglial cells through inserts with 8 µm pores. ( A ) The migration rate of primary microglial cells after the spared nerve injury (SNI) surgery is reduced compared with sham conditions. Histograms showing the contribution of each potassium channel, Kv1.3, Kv1.5 and Kir2.1, to microglial migration, in both sham ( B – D ) and SNI conditions ( E – G ). All the statistical analysis is represented as Mean ± SD, *: p

    Journal: International Journal of Molecular Sciences

    Article Title: Potassium Channels Kv1.3 and Kir2.1 But Not Kv1.5 Contribute to BV2 Cell Line and Primary Microglial Migration

    doi: 10.3390/ijms22042081

    Figure Lengend Snippet: The migration of primary microglial cells through inserts with 8 µm pores. ( A ) The migration rate of primary microglial cells after the spared nerve injury (SNI) surgery is reduced compared with sham conditions. Histograms showing the contribution of each potassium channel, Kv1.3, Kv1.5 and Kir2.1, to microglial migration, in both sham ( B – D ) and SNI conditions ( E – G ). All the statistical analysis is represented as Mean ± SD, *: p

    Article Snippet: The potassium channels were visualized with primary antibodies anti-Kv1.3, anti-Kv1.5, anti-Kir2.1 (#APC-002, #APC-004, #APC-0026 Alomone, Jerusalem, Israel), used in a dilution of 1:300 and the secondary antibody was goat anti-rabbit Alexa Fluor 568 (1:1500, #A11011, Life Technologies, Carlsbad, CA, USA).

    Techniques: Migration

    Decrease of P2Y6 Receptor and Calcium mobilization after treatment without change in intracellular PAD4 levels. (A) Neutrophils were treated as described before. P2Y6 was measured by flow cytometry with a FITC-conjugated antibody. Treatment with Alveofact ® decreases extracellular expression of the P2Y6 receptor independent of timing or dosing, while Curosurf ® does not exhibit this effect in every dosing. (B) Calcium levels at 3 h after treatment were used. Data were normalized to stimulated PMA control due to the heterogeneity in basic calcium levels and the response of the neutrophils to stimulation. Results are similar to the findings of the P2Y6 receptor expression. (C) Intracellular PAD4 levels were determined by ELISA using cell lysates of 1×10 6 cells. Interestingly, PAD4 levels did not change after treatment with surfactant compared to PMA-stimulated neutrophils. Significance level was set as p

    Journal: Frontiers in Immunology

    Article Title: The Inhibitory Effect of Curosurf® and Alveofact® on the Formation of Neutrophil Extracellular Traps

    doi: 10.3389/fimmu.2020.582895

    Figure Lengend Snippet: Decrease of P2Y6 Receptor and Calcium mobilization after treatment without change in intracellular PAD4 levels. (A) Neutrophils were treated as described before. P2Y6 was measured by flow cytometry with a FITC-conjugated antibody. Treatment with Alveofact ® decreases extracellular expression of the P2Y6 receptor independent of timing or dosing, while Curosurf ® does not exhibit this effect in every dosing. (B) Calcium levels at 3 h after treatment were used. Data were normalized to stimulated PMA control due to the heterogeneity in basic calcium levels and the response of the neutrophils to stimulation. Results are similar to the findings of the P2Y6 receptor expression. (C) Intracellular PAD4 levels were determined by ELISA using cell lysates of 1×10 6 cells. Interestingly, PAD4 levels did not change after treatment with surfactant compared to PMA-stimulated neutrophils. Significance level was set as p

    Article Snippet: Cells were washed twice with PBS (Thermo Fischer, Waltham, MA, USA) and labeled with propidium iodide (PI) and Annexin-V-FITC (Becton, Dickinson and Company, Franklin Lakes, NJ, USA) or anti-CD11b-VioBlue (mAb REA713, IgG1) and anti-CD66b-APC (mAb REA306, IgG1) antibodies (Miltenyi Biotec, Bergisch Gladbach, Germany) or anti-P2Y6 Receptor-FITC (Alomone Labs, Jerusalem, Israel) as described in the manufacturer’s protocols.

    Techniques: Flow Cytometry, Expressing, Enzyme-linked Immunosorbent Assay