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Alomone Labs rabbit anti scn5a
Figure 1. The intracellular level of NADH is increased by A280V GPD1-L and is reversed by incubation with [NAD]o. Data from 3 to 4 samples are normalized to the <t>SCN5A</t> cell group. ***P0.001 vs all other groups.

Figure 1. The intracellular level of NADH is increased by A280V GPD1-L and is reversed by incubation with [NAD]o. Data from 3 to 4 samples are normalized to the <t>SCN5A</t> cell group. ***P0.001 vs all other groups.

Rabbit Anti Scn5a, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 79 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/asc-045/Anti-NaV1%2E5+Antibody/10__1161_slash_circresaha__109__197277-186-40-43
Average 93 stars, based on 79 article reviews

rabbit anti scn5a - by Bioz Stars, 2026-10

93/100 stars

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1) Product Images from "Cardiac Na + Current Regulation by Pyridine Nucleotides"

Article Title: Cardiac Na + Current Regulation by Pyridine Nucleotides

Journal: Circulation Research

doi: 10.1161/circresaha.109.197277

Figure Legend Snippet: Figure 1. The intracellular level of NADH is increased by A280V GPD1-L and is reversed by incubation with [NAD]o. Data from 3 to 4 samples are normalized to the SCN5A cell group. ***P0.001 vs all other groups.

Techniques Used: Incubation

Figure 5. The effects of altering NAD(H) on arrhythmic risk. Representative MAP recordings with PES during Krebs–Henseleit buffer per- fusion (A), after 20 minutes perfusion with lactate:pyruvate (10:1) buffer to increase [NADH]i (B), and after reintroduction of Krebs–H- enseleit buffer (C). D, Representative traces of MAPs from left ventricular epicardium of Langendorff-perfused SCN5A/ heart during standard pacing at a basic cycle length of 125 ms in the control condition. Vertical lines below the MAPs represent the times when electric stimulations were delivered. E, Action potentials recorded with the patch-clamp technique in single ventricular myocytes and with the MAP electrode on whole heart. F, MAPs after 20 minutes of perfusion with 100 mol/L [NAD]o. G, Histogram of APD90 in control condition and with 100 mol/L [NAD]o. H, Representative MAPs recorded during PES showing PES-induced VT in SCN5A/

Figure Legend Snippet: Figure 5. The effects of altering NAD(H) on arrhythmic risk. Representative MAP recordings with PES during Krebs–Henseleit buffer per- fusion (A), after 20 minutes perfusion with lactate:pyruvate (10:1) buffer to increase [NADH]i (B), and after reintroduction of Krebs–H- enseleit buffer (C). D, Representative traces of MAPs from left ventricular epicardium of Langendorff-perfused SCN5A/ heart during standard pacing at a basic cycle length of 125 ms in the control condition. Vertical lines below the MAPs represent the times when electric stimulations were delivered. E, Action potentials recorded with the patch-clamp technique in single ventricular myocytes and with the MAP electrode on whole heart. F, MAPs after 20 minutes of perfusion with 100 mol/L [NAD]o. G, Histogram of APD90 in control condition and with 100 mol/L [NAD]o. H, Representative MAPs recorded during PES showing PES-induced VT in SCN5A/

Techniques Used: Control, Patch Clamp

Figure Legend Snippet: Figure 6. Possible signaling pathways for the effects of A280V GPD1-L, [NADH]i and NAD to regulate the Nav1.5.

Techniques Used: Protein-Protein interactions

Western Blot:

Article Title: Popeye domain containing proteins modulate the voltage-gated cardiac sodium channel Nav1.5
Article Snippet: For CoIP Dynabeads protein A magnetic beads (Thermo Fisher Scientific) were used following manufacturer’s instructions. .. Anti-Nav1.5 (SCN5A) antibody (493-511, Alomone) was used for precipitation and Western Blots were stained with anti-Nav1.5 (SCN5A) (493-511, Alomone, 1:1000) or anti-POPDC2 (Sigma, HPA 024255, 1:500) antibodies and peroxidase-conjugated anti-rabbit secondary antibody (Thermo, 31460, 1:8000). ..

Staining:

Labeling:

Article Title: Cardiac Na + Current Regulation by Pyridine Nucleotides
Article Snippet: .. For biotin labeling, SCN5A cells were incubated as above for 2-10 min. Biotinylation of cell surface proteins was performed with the Pinpoint Cell Surface Protein Isolation Kit (Pierce, Rockford, IL) as previously described.3 For detection of SCN5A, the primary antibody (rabbit anti- SCN5A, Alomone Labs, Jerusalem, Israel) was diluted 1:100. .. Alkaline phosphatase-conjugated goat anti-rabbit IgG secondary antibody (Jackson ImmunoResearch, West Grove, PA, USA) was diluted 1:7500.

Incubation:

Article Title: Candesartan Cilexetil Attenuates Arrhythmogenicity Following Pressure Overload in Rats via the Modulation of Cardiac Electrical and Structural Remodeling and Calcium Handling Dysfunction
Article Snippet: Protein samples (30– 100 μg) were separated on 4% to 20% gradient gel (Mini- Protean Precast gels; Bio- Rad Laboratories) or 8% to 10% SDS- PAGE and transferred onto polyvinylidene difluoride (PVDF) membranes (PerkinElmer Life Sciences, Boston, MA). .. Membranes were blocked and then incubated overnight at 4 °C with primary antibody against ryanodine receptor Ca2+ release channel 2 (RyR2) (1:1000; ThermoFisher Scientific, Waltham, MA), Ser2808- phosphorylated RyR2 (pSer2808- RyR2) (1:5000; Badrilla, Leeds, United Kingdom), pSer2814- RyR2 (1:500; Badrilla), SERCA2a (sarco[endo]plasmic reticulum Ca2+ATPase) (1:5000; Badrilla), phospholamban (1:2000; Badrilla), pSer16- phospholamban (1:2000; Millipore, Temecula, CA), pThr17- phospholamban (1:2000; Badrilla), Na+- Ca2+ exchanger 1 (NCX1) (1:500; Santa Cruz Biotechnology, Santa Cruz, CA), voltage- gated L- type calcium 1.2a channel subunit (CaV1.2a) (1:400; Millipore), voltage- gated sodium 1.5 channel subunit (NaV1.5) (1:500; Alomone, Jerusalem, Israel), voltagegated potassium 4.2 channel subunit (Kv4.2) (1:200; Millipore), voltage- gated potassium 4.3 channel subunit (Kv4.3) (1:200; Alomone), voltage- gated potassium 1.4 channel subunit (Kv1.4) (1:200; Alomone), Kv- channel interacting protein 2 (KChIP2) (1:500; ThermoFisher Scientific), voltage- gated potassium 2.1 channel subunit (Kv2.1) (1:500; Alomone), and inwardly rectifying potassium 2.1 channel subunit (Kir2.1) (1:600; Alomone). .. The blot was also probed by a glyceraldehyde 3- phosphate dehydrogenase (GAPDH) antibody (1:1000; Santa Cruz) or a β- actin antibody (1:1000; Abcam, Cambridge, MA) as an internal control.

Isolation:

Article Title: Cardiac Na + Current Regulation by Pyridine Nucleotides
Article Snippet: .. Biotinylation of cell surface proteins was performed with the Pinpoint Cell Surface Protein Isolation Kit (Pierce, Rockford, Ill) as previously described.7 For detection of SCN5A, the primary antibody (rabbit anti-SCN5A, Alomone Labs, Jerusalem, Israel) was diluted 1:100. .. For imaging, HEK293 cells were transfected with SCN5A-GFP cDNA, in which the cDNA sequence encoding GFP was fused to the C terminus of SCN5A.18 Cells were incubated with lactate:pyruvate (10 mmol/ L:1 mmol/L) to raise intracellular NADH20–22 or treated with phorbol 12-myristate 13-acetate (PMA) (30 nmol/L) for 10 minutes and then fixed for fluorescent imaging with a Zeiss LSM510 META confocal microscope (Carl Zeiss, Jena, Germany).

Membrane:

Article Title: Single-Molecule Localization of the Cardiac Voltage-Gated Sodium Channel Reveals Different Modes of Reorganization at Cardiomyocyte Membrane Domains
Article Snippet: Secondary antibodies were rabbit AlexaFluor 568 (Life Technologies, Carlsbad, California; SMLM: 1:10,000; confocal: 1:200), mouse AlexaFluor 647 (Life Technologies; SMLM: 1:5000), and mouse AlexaFluor 488 (Life Technologies; confocal: 1:200). .. Our group has shown a similar reduction of Nav1.5 at the lateral membrane of mdx mice using the aforementioned Alomone antibody9. .. Firstly, the confocal image in Figure 1 shows that the staining pattern of the Pineda antibody is highly similar to that of the widely used Alomone ASC-005 antibody2.

Image Search Results

Functional screen for cardiac ion channels modulated by POPDC2 (A–D) Representative current traces of (A) HCN4, applying a voltage step from −30 mV to −120 mV within 6 s, from a holding potential of −30 mV, (B) Kv1.4, applying a voltage step from −80 mV to +40 mV within 2 s, from a holding potential of −80 mV, (C) Kv4.3, applying a voltage step from −80 mV to +40 mV within 2 s, from a holding potential of −80 mV or (D) Nav1.5, applying a voltage step from −80 mV to −20 mV within 50 ms, from a holding potential of −80 mV, expressed alone ( black ) or together with POPDC2 ( blue ) in Xenopus laevis oocytes. (E) Indicated cardiac channels or the Na + /K + ATPase (Na/K) were expressed alone or together with POPDC2 in Xenopus oocytes and relative changes in current amplitudes by POPDC2 were plotted. Oocytes were stored in a storage solution lacking theophylline. The number of technical replicates (n) is indicated in parentheses. Data are presented as mean ± s.e.m. Significance was probed using a two-sided Student’s t-test. ∗∗∗, p < 0.001.

Journal: iScience

Article Title: Popeye domain containing proteins modulate the voltage-gated cardiac sodium channel Nav1.5

doi: 10.1016/j.isci.2024.109696

Figure Lengend Snippet: Functional screen for cardiac ion channels modulated by POPDC2 (A–D) Representative current traces of (A) HCN4, applying a voltage step from −30 mV to −120 mV within 6 s, from a holding potential of −30 mV, (B) Kv1.4, applying a voltage step from −80 mV to +40 mV within 2 s, from a holding potential of −80 mV, (C) Kv4.3, applying a voltage step from −80 mV to +40 mV within 2 s, from a holding potential of −80 mV or (D) Nav1.5, applying a voltage step from −80 mV to −20 mV within 50 ms, from a holding potential of −80 mV, expressed alone ( black ) or together with POPDC2 ( blue ) in Xenopus laevis oocytes. (E) Indicated cardiac channels or the Na + /K + ATPase (Na/K) were expressed alone or together with POPDC2 in Xenopus oocytes and relative changes in current amplitudes by POPDC2 were plotted. Oocytes were stored in a storage solution lacking theophylline. The number of technical replicates (n) is indicated in parentheses. Data are presented as mean ± s.e.m. Significance was probed using a two-sided Student’s t-test. ∗∗∗, p < 0.001.

Article Snippet: Anti-Nav1.5 (SCN5A) antibody (493-511, Alomone) was used for precipitation and Western Blots were stained with anti-Nav1.5 (SCN5A) (493-511, Alomone, 1:1000) or anti-POPDC2 (Sigma, HPA 024255, 1:500) antibodies and peroxidase-conjugated anti-rabbit secondary antibody (Thermo, 31460, 1:8000).

Techniques: Functional Assay, Patch Clamp

Interaction of POPDC proteins with the cardiac sodium channel Nav1.5 or the skeletal muscle sodium channel Nav1.4 (A) Co-immunoprecipitation of POPDC2-FLAG with Nav1.5-HA was performed in COS-7 cells using anti-HA antibodies. An empty vector served as control. The co-immunoprecipitation was subsequently detected with an anti-FLAG antibody (IB: α-Flag). (B) Co-immunoprecipitation of POPDC2 and Nav1.5 in murine heart lysates (mheart) using anti Nav1.5 antibodies (IP: α-Nav1.5). The immunoprecipitation was subsequently detected with an anti-Nav1.5 (IB: α-Nav1.5) and the co-immunoprecipitation by an anti-POPDC2 antibody (IB: α-POPDC2). HeLa cells were used as a control. (C) Oocytes were injected with cRNA for human Nav1.5 or co-injected with different murine POPDC isoforms and subsequently stored in a storage solution lacking theophylline. Currents were recorded with a P/N ( N = 4) voltage-step protocol (holding potential −80 mV) and voltage-steps of 50 ms ranging from −70 to +80 mV in 10 mV increments (see inset). (D and E) (D) Relative current amplitudes analyzed at −20 mV and normalized to Nav1.5. (E) Voltage-dependence of Nav1.5 activation in the absence or presence of POPDC1, POPDC2 or POPDC3. The respective voltages of half-maximal activation (V 1/2 ) and the Boltzmann constants (k) are provided as inset. The number of technical replicates (n) is indicated in parentheses or bar graphs. Data are presented as mean ± s.e.m. Significance was probed using a two-sided Student’s t-test. ∗∗∗, p < 0.001. (F and G) (F) POPDC2 does not modulate current amplitudes of Nav1.4, analyzed at −10 mV and normalized to Nav1.4 or (G) the shape of the voltage-dependence of Nav1.4 activation. The respective voltages of half-maximal activation (V 1/2 ) and the Boltzmann constants (k) are provided as inset. The number of technical replicates (n) is indicated in parentheses or bar graphs. Oocytes were stored in a storage solution lacking theophylline. Data are presented as mean ± s.e.m. Significance was probed using a two-sided Student’s t-test. See also <xref ref-type=

Figures S1 and . " width="100%" height="100%">

Journal: iScience

Article Title: Popeye domain containing proteins modulate the voltage-gated cardiac sodium channel Nav1.5

doi: 10.1016/j.isci.2024.109696

Figure Lengend Snippet: Interaction of POPDC proteins with the cardiac sodium channel Nav1.5 or the skeletal muscle sodium channel Nav1.4 (A) Co-immunoprecipitation of POPDC2-FLAG with Nav1.5-HA was performed in COS-7 cells using anti-HA antibodies. An empty vector served as control. The co-immunoprecipitation was subsequently detected with an anti-FLAG antibody (IB: α-Flag). (B) Co-immunoprecipitation of POPDC2 and Nav1.5 in murine heart lysates (mheart) using anti Nav1.5 antibodies (IP: α-Nav1.5). The immunoprecipitation was subsequently detected with an anti-Nav1.5 (IB: α-Nav1.5) and the co-immunoprecipitation by an anti-POPDC2 antibody (IB: α-POPDC2). HeLa cells were used as a control. (C) Oocytes were injected with cRNA for human Nav1.5 or co-injected with different murine POPDC isoforms and subsequently stored in a storage solution lacking theophylline. Currents were recorded with a P/N ( N = 4) voltage-step protocol (holding potential −80 mV) and voltage-steps of 50 ms ranging from −70 to +80 mV in 10 mV increments (see inset). (D and E) (D) Relative current amplitudes analyzed at −20 mV and normalized to Nav1.5. (E) Voltage-dependence of Nav1.5 activation in the absence or presence of POPDC1, POPDC2 or POPDC3. The respective voltages of half-maximal activation (V 1/2 ) and the Boltzmann constants (k) are provided as inset. The number of technical replicates (n) is indicated in parentheses or bar graphs. Data are presented as mean ± s.e.m. Significance was probed using a two-sided Student’s t-test. ∗∗∗, p < 0.001. (F and G) (F) POPDC2 does not modulate current amplitudes of Nav1.4, analyzed at −10 mV and normalized to Nav1.4 or (G) the shape of the voltage-dependence of Nav1.4 activation. The respective voltages of half-maximal activation (V 1/2 ) and the Boltzmann constants (k) are provided as inset. The number of technical replicates (n) is indicated in parentheses or bar graphs. Oocytes were stored in a storage solution lacking theophylline. Data are presented as mean ± s.e.m. Significance was probed using a two-sided Student’s t-test. See also Figures S1 and .

Techniques: Co-Immunoprecipitation Assay, Plasmid Preparation, Control, Immunoprecipitation, Injection, Activation Assay, Patch Clamp

POPDC2 does not alter the current kinetics of Nav1.5 (A) Current traces of human Nav1.5 expressed alone or co-expressed with human POPDC2. Representative traces are depicted at −30 mV, −20 mV and −10 mV. (B) Voltage-dependence of activation and inactivation curves fitted to a Boltzmann equation. The respective voltages of half-maximal activation (V 1/2 ) together with the Boltzmann constants (k), as well as the window currents, are provided as inset. For the V 1/2 of activation measurements, the currents were recorded with a P/N ( N = 4) voltage-step protocol (holding potential −80 mV) and voltage-steps of 50 ms ranging from −70 to +80 mV in 10 mV increments (see inset activation protocol). For the V 1/2 of inactivation measurements, the currents were recorded with a P/N ( N = 4) voltage-step protocol (holding potential −100 mV) and voltage-steps of 50 ms ranging from −120 to −10 mV in 10 mV increments with a final step to −30 mV for 50 ms (see inset inactivation protocol). The number of technical replicates (n) is indicated in parentheses. Data are presented as mean ± s.e.m. (C) Representative current traces of the kinetics of inactivation recorded with a single-step protocol from −100 mV to −30 mV for 65 ms (inset illustrates voltage protocol). (D) Representative recordings of the recovery from inactivation (inset illustrates voltage protocol). (E) Time-course of the recovery from inactivation recorded with a two-pulse protocol, first step to −10 mV for 50 ms and the second test pulse stepped to −10 mV with increasing interpulse intervals. The number of technical replicates (n) is indicated in parentheses. Data are presented as mean ± s.e.m. (F) Time constants of the fast and slow component of the recovery from inactivation (lower panel) and the relative contribution of the fast and slow component (upper panel). Values were obtained by a bi-exponential fit from the data in (E). Oocytes were stored in a storage solution lacking theophylline. The number of technical replicates (n) is indicated in parentheses. Data are presented as mean ± s.e.m. See also <xref ref-type=

Figure S3 . " width="100%" height="100%">

Journal: iScience

Article Title: Popeye domain containing proteins modulate the voltage-gated cardiac sodium channel Nav1.5

doi: 10.1016/j.isci.2024.109696

Figure Lengend Snippet: POPDC2 does not alter the current kinetics of Nav1.5 (A) Current traces of human Nav1.5 expressed alone or co-expressed with human POPDC2. Representative traces are depicted at −30 mV, −20 mV and −10 mV. (B) Voltage-dependence of activation and inactivation curves fitted to a Boltzmann equation. The respective voltages of half-maximal activation (V 1/2 ) together with the Boltzmann constants (k), as well as the window currents, are provided as inset. For the V 1/2 of activation measurements, the currents were recorded with a P/N ( N = 4) voltage-step protocol (holding potential −80 mV) and voltage-steps of 50 ms ranging from −70 to +80 mV in 10 mV increments (see inset activation protocol). For the V 1/2 of inactivation measurements, the currents were recorded with a P/N ( N = 4) voltage-step protocol (holding potential −100 mV) and voltage-steps of 50 ms ranging from −120 to −10 mV in 10 mV increments with a final step to −30 mV for 50 ms (see inset inactivation protocol). The number of technical replicates (n) is indicated in parentheses. Data are presented as mean ± s.e.m. (C) Representative current traces of the kinetics of inactivation recorded with a single-step protocol from −100 mV to −30 mV for 65 ms (inset illustrates voltage protocol). (D) Representative recordings of the recovery from inactivation (inset illustrates voltage protocol). (E) Time-course of the recovery from inactivation recorded with a two-pulse protocol, first step to −10 mV for 50 ms and the second test pulse stepped to −10 mV with increasing interpulse intervals. The number of technical replicates (n) is indicated in parentheses. Data are presented as mean ± s.e.m. (F) Time constants of the fast and slow component of the recovery from inactivation (lower panel) and the relative contribution of the fast and slow component (upper panel). Values were obtained by a bi-exponential fit from the data in (E). Oocytes were stored in a storage solution lacking theophylline. The number of technical replicates (n) is indicated in parentheses. Data are presented as mean ± s.e.m. See also Figure S3 .

Techniques: Activation Assay, Inactivation, Patch Clamp

POPDC2 reduces Nav1.5 current amplitudes in a cAMP-dependent manner (A) POPDC2 modulation of Nav1.5 is cAMP-dependent. Representative current traces recorded by voltage steps to −20 mV. Oocytes were injected with 10 ng Nav1.5 alone or 10 ng Nav1.5 together with 5 ng POPDC2 (50%) or 10 ng POPDC2 (100%), and stored in ND96 solution with or without theophylline before voltage-clamp measurements in oocytes. Inset illustrates the applied voltage protocol. (B) Analyses of peak current amplitudes from experiments performed in (A), analyzed at −20 mV and normalized to Nav1.5. The number of technical replicates (n) is indicated in bar graphs. Data are presented as mean ± s.e.m. Significance was probed using a two-sided Student’s t-test. ∗, p < 0.05; ∗∗∗, p < 0.001. (C) Oocytes were injected as in (A-B) but incubated in a storage solution containing theophylline. Relative current amplitudes were analyzed at −20 mV and normalized to Nav1.5. The number of technical replicates is indicated in bar graphs. Data are presented as mean ± s.e.m. See also <xref ref-type=

Figure S4 . " width="100%" height="100%">

Journal: iScience

Article Title: Popeye domain containing proteins modulate the voltage-gated cardiac sodium channel Nav1.5

doi: 10.1016/j.isci.2024.109696

Figure Lengend Snippet: POPDC2 reduces Nav1.5 current amplitudes in a cAMP-dependent manner (A) POPDC2 modulation of Nav1.5 is cAMP-dependent. Representative current traces recorded by voltage steps to −20 mV. Oocytes were injected with 10 ng Nav1.5 alone or 10 ng Nav1.5 together with 5 ng POPDC2 (50%) or 10 ng POPDC2 (100%), and stored in ND96 solution with or without theophylline before voltage-clamp measurements in oocytes. Inset illustrates the applied voltage protocol. (B) Analyses of peak current amplitudes from experiments performed in (A), analyzed at −20 mV and normalized to Nav1.5. The number of technical replicates (n) is indicated in bar graphs. Data are presented as mean ± s.e.m. Significance was probed using a two-sided Student’s t-test. ∗, p < 0.05; ∗∗∗, p < 0.001. (C) Oocytes were injected as in (A-B) but incubated in a storage solution containing theophylline. Relative current amplitudes were analyzed at −20 mV and normalized to Nav1.5. The number of technical replicates is indicated in bar graphs. Data are presented as mean ± s.e.m. See also Figure S4 .

Techniques: Injection, Incubation, Patch Clamp

POPDC2 co-localizes with Nav1.5 at the plasma membrane and intracellular compartments and regulates the surface expression of Nav1.5 in a cAMP-dependent manner (A and C) HeLa cells were transfected exclusively with Nav1.5-HA or (B and D) in combination with POPDC2-YFP. 24 h after transfection the cells were fixed, non-permeabilized (A-B) or permeabilized (C-D) and immunostained with anti HA antibody and Alexa 647 conjugated secondary antibody. The cells were then analyzed by confocal microscopy. Nuclear counterstaining with Hoechst 33342 is depicted in blue and nuclei of non-transfected cells are indicated by asterisk. Arrows point to either (A) Nav1.5- or (B) Nav1.5- plus POPDC2-YFP-stained membrane protrusions; PM, plasma membrane. Arrowheads in (D) depict Nav1.5-positive intracellular structures co-labelled by POPDC2-YFP, which are also designated by arrows in the single fluorescence images shown on the left. Scale bars, 10 μm. (E) Cartoon of the chemiluminescence-based ELISA-assay performed in HeLa cells transiently transfected with Nav1.5 HA to quantify the changes in Nav1.5 surface expression by co-transfection with POPDC2. In fixed cells, the primary antibody detects extracellularly HA-tagged Nav1.5 channels (Nav1.5 HA ) at the surface membrane while the peroxidase-enzyme (POD) which is linked to the secondary antibody catalyzes the oxidation of a luminol-substrate which is accompanied by the emission of light. Subsequently, the light emission can be quantified as relative light units (RLUs) as a measure of the surface expression of the channel. (F) Chemiluminescence assay of extracellularly HA-tagged Nav1.5 to determine the relative changes in surface expression by co-expressed POPDC2. The number of biological replicates (N) is indicated in the bar graphs. Data are expressed as relative light units (RLUs). Data are presented as mean ± s.e.m. Significance was probed using a two-sided Student’s t-test. ∗, p < 0.05. (G) Chemiluminescence assay of extracellularly HA-tagged Nav1.5 to determine the relative changes in surface expression by co-expressed POPDC2 in the presence of theophylline. The number of biological replicates (N) is indicated in the bar graphs. Data are expressed as relative light units (RLUs). Data are presented as mean ± s.e.m.

Journal: iScience

Article Title: Popeye domain containing proteins modulate the voltage-gated cardiac sodium channel Nav1.5

doi: 10.1016/j.isci.2024.109696

Figure Lengend Snippet: POPDC2 co-localizes with Nav1.5 at the plasma membrane and intracellular compartments and regulates the surface expression of Nav1.5 in a cAMP-dependent manner (A and C) HeLa cells were transfected exclusively with Nav1.5-HA or (B and D) in combination with POPDC2-YFP. 24 h after transfection the cells were fixed, non-permeabilized (A-B) or permeabilized (C-D) and immunostained with anti HA antibody and Alexa 647 conjugated secondary antibody. The cells were then analyzed by confocal microscopy. Nuclear counterstaining with Hoechst 33342 is depicted in blue and nuclei of non-transfected cells are indicated by asterisk. Arrows point to either (A) Nav1.5- or (B) Nav1.5- plus POPDC2-YFP-stained membrane protrusions; PM, plasma membrane. Arrowheads in (D) depict Nav1.5-positive intracellular structures co-labelled by POPDC2-YFP, which are also designated by arrows in the single fluorescence images shown on the left. Scale bars, 10 μm. (E) Cartoon of the chemiluminescence-based ELISA-assay performed in HeLa cells transiently transfected with Nav1.5 HA to quantify the changes in Nav1.5 surface expression by co-transfection with POPDC2. In fixed cells, the primary antibody detects extracellularly HA-tagged Nav1.5 channels (Nav1.5 HA ) at the surface membrane while the peroxidase-enzyme (POD) which is linked to the secondary antibody catalyzes the oxidation of a luminol-substrate which is accompanied by the emission of light. Subsequently, the light emission can be quantified as relative light units (RLUs) as a measure of the surface expression of the channel. (F) Chemiluminescence assay of extracellularly HA-tagged Nav1.5 to determine the relative changes in surface expression by co-expressed POPDC2. The number of biological replicates (N) is indicated in the bar graphs. Data are expressed as relative light units (RLUs). Data are presented as mean ± s.e.m. Significance was probed using a two-sided Student’s t-test. ∗, p < 0.05. (G) Chemiluminescence assay of extracellularly HA-tagged Nav1.5 to determine the relative changes in surface expression by co-expressed POPDC2 in the presence of theophylline. The number of biological replicates (N) is indicated in the bar graphs. Data are expressed as relative light units (RLUs). Data are presented as mean ± s.e.m.

Techniques: Clinical Proteomics, Membrane, Expressing, Transfection, Confocal, Microscopy, Staining, Fluorescence, Enzyme-linked Immunosorbent Assay, Cotransfection, Chemiluminescence Immunoassay, Immunofluorescence

Journal: iScience

Article Title: Popeye domain containing proteins modulate the voltage-gated cardiac sodium channel Nav1.5

doi: 10.1016/j.isci.2024.109696

Figure Lengend Snippet:

Techniques: Recombinant, Magnetic Beads, Enzyme-linked Immunosorbent Assay, Software, Spectrophotometry, Confocal, Microscopy

Journal: iScience

Article Title: Popeye domain containing proteins modulate the voltage-gated cardiac sodium channel Nav1.5

doi: 10.1016/j.isci.2024.109696

Figure Lengend Snippet:

Techniques: Patch Clamp

Journal: Circulation Research

Article Title: Cardiac Na + Current Regulation by Pyridine Nucleotides

doi: 10.1161/circresaha.109.197277

Figure Lengend Snippet: Figure 1. The intracellular level of NADH is increased by A280V GPD1-L and is reversed by incubation with [NAD]o. Data from 3 to 4 samples are normalized to the SCN5A cell group. ***P0.001 vs all other groups.

Article Snippet: For biotin labeling, SCN5A cells were incubated as above for 2-10 min. Biotinylation of cell surface proteins was performed with the Pinpoint Cell Surface Protein Isolation Kit (Pierce, Rockford, IL) as previously described.3 For detection of SCN5A, the primary antibody (rabbit anti- SCN5A, Alomone Labs, Jerusalem, Israel) was diluted 1:100.

Techniques: Incubation

Journal: Circulation Research

Article Title: Cardiac Na + Current Regulation by Pyridine Nucleotides

doi: 10.1161/circresaha.109.197277

Figure Lengend Snippet: Figure 5. The effects of altering NAD(H) on arrhythmic risk. Representative MAP recordings with PES during Krebs–Henseleit buffer per- fusion (A), after 20 minutes perfusion with lactate:pyruvate (10:1) buffer to increase [NADH]i (B), and after reintroduction of Krebs–H- enseleit buffer (C). D, Representative traces of MAPs from left ventricular epicardium of Langendorff-perfused SCN5A/ heart during standard pacing at a basic cycle length of 125 ms in the control condition. Vertical lines below the MAPs represent the times when electric stimulations were delivered. E, Action potentials recorded with the patch-clamp technique in single ventricular myocytes and with the MAP electrode on whole heart. F, MAPs after 20 minutes of perfusion with 100 mol/L [NAD]o. G, Histogram of APD90 in control condition and with 100 mol/L [NAD]o. H, Representative MAPs recorded during PES showing PES-induced VT in SCN5A/

Techniques: Control, Patch Clamp

Journal: Circulation Research

Article Title: Cardiac Na + Current Regulation by Pyridine Nucleotides

doi: 10.1161/circresaha.109.197277

Figure Lengend Snippet: Figure 6. Possible signaling pathways for the effects of A280V GPD1-L, [NADH]i and NAD to regulate the Nav1.5.

Techniques: Protein-Protein interactions

Review

rabbit anti scn5a (Alomone Labs)

Structured Review

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1) Product Images from "Cardiac Na + Current Regulation by Pyridine Nucleotides"

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