l glutamine  (Biochrom)

 
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    Biochrom l glutamine
    L Glutamine, supplied by Biochrom, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/l glutamine/product/Biochrom
    Average 86 stars, based on 1 article reviews
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    Cell Culture:

    Article Title: Cold Atmospheric Plasma Induces Apoptosis and Oxidative Stress Pathway Regulation in T-Lymphoblastoid Leukemia Cells
    Article Snippet: We demonstrated that the exposure of complete medium to CAP produced by a nanosecond-pulsed DBD induced the generation of several RONS; among these species, nitrites and hydrogen peroxide are considered the most significant RONS contributing to plasma toxicity on cancer cells. .. Cell Culture Jurkat cells were purchased from LGC Standards (Teddington, UK) and cultured in RPMI 1640 supplemented with 10% heat-inactivated fetal calf serum, 1% antibiotics [penicillin 5000 IU/streptomycin 5 mg/mL], and 1% L-glutamine solution (all purchased from Biochrom, Billerica, MA, USA). ..

    Article Title: Isolation, Identification, and Characterization of Novel Arenaviruses, the Etiological Agents of Boid Inclusion Body Disease
    Article Snippet: Supernatants were centrifuged (500 × g at room temperature [RT]) for 5 min, and cells were suspended in 5 ml of HEPES buffered cell culture medium with 10% fetal bovine serum (FBS; Biochrom), inactivated at 56°C for 30 min in sterile cell culture dishes 5 cm in diameter, and incubated at 30°C. .. One liter of cell culture medium was prepared containing 873.5 ml of basal medium Eagle, (1× BME; Biochrom, Berlin, Germany) with 100 ml of tryptose phosphate broth (TPB [Difco, Sigma-Aldrich, Germany]; 29.5 g solubilized in 1 liter of aqua bidest, autoclaved at 121°C for 21 min), 15 ml of HEPES buffer (1 M; Biochrom), 10 ml of l -glutamine (200 mM l -glutamine; Biochrom), 1 ml of gentamicin (10 mg/ml; Biochrom), and 0.5 ml of nystatin (100,000 IU/ml Nystatin Lederle; Valeant Pharmaceuticals, Eschborn, Germany), pH 7.2 to 7.3. ..

    Article Title: Macrophage migration inhibitory factor (MIF) plays a pivotal role in immunity against Salmonella typhimurium
    Article Snippet: .. Spleen cells (2 × 105 per well) were cultured in 96-well, round-bottomed plates in RPMI medium 1640 supplemented with 2 mM l -glutamine/1 mM Na-pyruvate/100 μg/ml penicillin/100 μg/ml streptomycin (all Biochrom, Berlin), 50 μM 2-mercaptoethanol (GIBCO/BRL), and 10% FCS (Sigma). ..

    Recombinant:

    Article Title: Leishmania disease development depends on the presence of apoptotic promastigotes in the virulent inoculum
    Article Snippet: PMN were isolated from buffy coat blood as described ( ). .. PMN (1 × 107 ml) were coincubated with stat. phase L. major promastigotes with or without preincubation of 5 μg of recombinant AnxA5 (Responsif, Erlangen, Germany) or with MACS-purified AnxA5+, AnxA5−, or a 1:1 mixture of AnxA5+ and AnxA5− parasites, at 37°C at a AnxA5− parasite/PMN ratio of 5:1 in RPMI medium 1640 (Gibco, Grand Island, NY), supplemented with 5% heat-inactivated FCS/50 μM 2-mercaptoethanol/2 mM l -glutamine/10 mM Hepes/100 μg/ml penicillin/160 μg/ml gentamicin (all from Seromed-Biochrom, Berlin, Germany). ..

    Magnetic Cell Separation:

    Article Title: Leishmania disease development depends on the presence of apoptotic promastigotes in the virulent inoculum
    Article Snippet: PMN were isolated from buffy coat blood as described ( ). .. PMN (1 × 107 ml) were coincubated with stat. phase L. major promastigotes with or without preincubation of 5 μg of recombinant AnxA5 (Responsif, Erlangen, Germany) or with MACS-purified AnxA5+, AnxA5−, or a 1:1 mixture of AnxA5+ and AnxA5− parasites, at 37°C at a AnxA5− parasite/PMN ratio of 5:1 in RPMI medium 1640 (Gibco, Grand Island, NY), supplemented with 5% heat-inactivated FCS/50 μM 2-mercaptoethanol/2 mM l -glutamine/10 mM Hepes/100 μg/ml penicillin/160 μg/ml gentamicin (all from Seromed-Biochrom, Berlin, Germany). ..

    Ex Vivo:

    Article Title: Modulation of the granzyme B inhibitor proteinase inhibitor 9 (PI-9) by activation of lymphocytes and monocytes in vitro and by Epstein-Barr virus and bacterial infection
    Article Snippet: For in-vitro cultivation, peripheral blood mononuclear cells (PBMC) were separated from fresh heparinzed blood of healthy adult donors by Biocoll (Biochrom, Berlin, Germany) density gradient centrifugation, followed by washing in phosphate-buffered saline (PBS, Biochrom). .. For ex-vivo incubation assays, cells were kept in RPMI-1640 medium (Life Technologies, Eggenstein, Germany) supplemented with 10% heat-inactivated fetal calf serum (FCS) (Conco, Wiesbaden, Germany), 12·5 mM HEPES (Biochrom, Berlin, Germany), 100 U/ml penicillin/streptomycin solution (Life Technologies) and 2·0 mM l -glutamine solution (Biochrom); they were seeded at a density of 1 × 106 cells/ml in six-well plates with the given stimuli or inhibitory substances for the given times at 37°C, 5% CO2 . ..

    Incubation:

    Article Title: Modulation of the granzyme B inhibitor proteinase inhibitor 9 (PI-9) by activation of lymphocytes and monocytes in vitro and by Epstein-Barr virus and bacterial infection
    Article Snippet: For in-vitro cultivation, peripheral blood mononuclear cells (PBMC) were separated from fresh heparinzed blood of healthy adult donors by Biocoll (Biochrom, Berlin, Germany) density gradient centrifugation, followed by washing in phosphate-buffered saline (PBS, Biochrom). .. For ex-vivo incubation assays, cells were kept in RPMI-1640 medium (Life Technologies, Eggenstein, Germany) supplemented with 10% heat-inactivated fetal calf serum (FCS) (Conco, Wiesbaden, Germany), 12·5 mM HEPES (Biochrom, Berlin, Germany), 100 U/ml penicillin/streptomycin solution (Life Technologies) and 2·0 mM l -glutamine solution (Biochrom); they were seeded at a density of 1 × 106 cells/ml in six-well plates with the given stimuli or inhibitory substances for the given times at 37°C, 5% CO2 . ..

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    • mmol ∙ l 1 glutamine  (Biochrom)
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    Biochrom mmol ∙ l 1 glutamine
    Effects of menthol and Mg 2+ on intracellular calcium [Ca 2+ ] i . a) Average of ten original recordings of control HEK-293 cells, showing a significant increase of Δ [Ca 2+ ] i in the 200 s interval after application of menthol (1 mmol∙l −1 ) (95% confidence interval in grey), with the timeline identical to that in b), showing the average [Ca 2+ ] i (± 95%) of eleven recordings of HEK-293 cells overexpressing bTRPV3. While resting levels of [Ca 2+ ] i in bTRPV3 cells were not significantly different from those of control cells, menthol led to a significantly higher Δ [Ca 2+ ] i in these cells (p = 0.003). c) Boxplots comparing the slopes (Δ [Ca 2+ ] i /Δ t) of the graphs in (a and b) in a 50 s interval at the beginning of the measurement and in the 50 s interval after application of menthol (differing letters above the boxes indicate p
    Mmol ∙ L 1 Glutamine, supplied by Biochrom, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mmol ∙ l 1 glutamine/product/Biochrom
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mmol ∙ l 1 glutamine - by Bioz Stars, 2021-03
    86/100 stars
    		  Buy from Supplier
    l glutamine  (Biochrom)
    86
    Biochrom l glutamine
    Effects of menthol and Mg 2+ on intracellular calcium [Ca 2+ ] i . a) Average of ten original recordings of control HEK-293 cells, showing a significant increase of Δ [Ca 2+ ] i in the 200 s interval after application of menthol (1 mmol∙l −1 ) (95% confidence interval in grey), with the timeline identical to that in b), showing the average [Ca 2+ ] i (± 95%) of eleven recordings of HEK-293 cells overexpressing bTRPV3. While resting levels of [Ca 2+ ] i in bTRPV3 cells were not significantly different from those of control cells, menthol led to a significantly higher Δ [Ca 2+ ] i in these cells (p = 0.003). c) Boxplots comparing the slopes (Δ [Ca 2+ ] i /Δ t) of the graphs in (a and b) in a 50 s interval at the beginning of the measurement and in the 50 s interval after application of menthol (differing letters above the boxes indicate p
    L Glutamine, supplied by Biochrom, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/l glutamine/product/Biochrom
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    l glutamine - by Bioz Stars, 2021-03
    86/100 stars
    		  Buy from Supplier

    Image Search Results


    Effects of menthol and Mg 2+ on intracellular calcium [Ca 2+ ] i . a) Average of ten original recordings of control HEK-293 cells, showing a significant increase of Δ [Ca 2+ ] i in the 200 s interval after application of menthol (1 mmol∙l −1 ) (95% confidence interval in grey), with the timeline identical to that in b), showing the average [Ca 2+ ] i (± 95%) of eleven recordings of HEK-293 cells overexpressing bTRPV3. While resting levels of [Ca 2+ ] i in bTRPV3 cells were not significantly different from those of control cells, menthol led to a significantly higher Δ [Ca 2+ ] i in these cells (p = 0.003). c) Boxplots comparing the slopes (Δ [Ca 2+ ] i /Δ t) of the graphs in (a and b) in a 50 s interval at the beginning of the measurement and in the 50 s interval after application of menthol (differing letters above the boxes indicate p

    Journal: PLoS ONE

    Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+

    doi: 10.1371/journal.pone.0193519

    Figure Lengend Snippet: Effects of menthol and Mg 2+ on intracellular calcium [Ca 2+ ] i . a) Average of ten original recordings of control HEK-293 cells, showing a significant increase of Δ [Ca 2+ ] i in the 200 s interval after application of menthol (1 mmol∙l −1 ) (95% confidence interval in grey), with the timeline identical to that in b), showing the average [Ca 2+ ] i (± 95%) of eleven recordings of HEK-293 cells overexpressing bTRPV3. While resting levels of [Ca 2+ ] i in bTRPV3 cells were not significantly different from those of control cells, menthol led to a significantly higher Δ [Ca 2+ ] i in these cells (p = 0.003). c) Boxplots comparing the slopes (Δ [Ca 2+ ] i /Δ t) of the graphs in (a and b) in a 50 s interval at the beginning of the measurement and in the 50 s interval after application of menthol (differing letters above the boxes indicate p

    Article Snippet: Cell culture and transient transfection of bTRPV3 HEK-293 cells were cultivated in Dulbecco’s modified Eagle’s medium supplemented with 10% (vol/vol) fetal bovine serum, 4 mmol∙l-1 glutamine and 100 units∙ml-1 of both penicillin and streptomycin (Biochrom, Berlin, Germany).

    Techniques:

    Original recordings of one patch from a bTRPV3 cell with NH 4 Cl in the pipette. a) Channel events are clearly visible in NaCl bath solution in response to pulse protocol III both at negative potentials and positive potentials, reflecting influx of Na + and efflux of NH 4 + . b) same patch in 72.5 mmol∙l −1 CaCl 2 bath solution. Ca 2+ had a clear blocking effect on the current level at positive potentials (efflux of NH 4 + ). c) Detail from (b), showing single-channel events at negative potentials, reflecting influx of Ca 2+ d) same patch in NMDGCl bath solution. e) Insert showing single-channel events at negative potential level, suggesting influx of NMDG + . f) Current-voltage plot of unitary current amplitudes from this individual patch, fitted with the current formulation of the Goldman-Hodgkin-Katz equation. The fit yields a conductance for Na + of 132 ± 4 pS, for Ca 2+ of 21 ± 3 pS, and for NMDG + of 36 ± 3 pS. The conductance for NH 4 + was similar in NaCl and NMDGCl solution (250 ± 3 pS or 264 ± 3 pS, respectively) but dropped to 88 ± 4 pS with high Ca 2+ in the bath.

    Journal: PLoS ONE

    Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+

    doi: 10.1371/journal.pone.0193519

    Figure Lengend Snippet: Original recordings of one patch from a bTRPV3 cell with NH 4 Cl in the pipette. a) Channel events are clearly visible in NaCl bath solution in response to pulse protocol III both at negative potentials and positive potentials, reflecting influx of Na + and efflux of NH 4 + . b) same patch in 72.5 mmol∙l −1 CaCl 2 bath solution. Ca 2+ had a clear blocking effect on the current level at positive potentials (efflux of NH 4 + ). c) Detail from (b), showing single-channel events at negative potentials, reflecting influx of Ca 2+ d) same patch in NMDGCl bath solution. e) Insert showing single-channel events at negative potential level, suggesting influx of NMDG + . f) Current-voltage plot of unitary current amplitudes from this individual patch, fitted with the current formulation of the Goldman-Hodgkin-Katz equation. The fit yields a conductance for Na + of 132 ± 4 pS, for Ca 2+ of 21 ± 3 pS, and for NMDG + of 36 ± 3 pS. The conductance for NH 4 + was similar in NaCl and NMDGCl solution (250 ± 3 pS or 264 ± 3 pS, respectively) but dropped to 88 ± 4 pS with high Ca 2+ in the bath.

    Article Snippet: Cell culture and transient transfection of bTRPV3 HEK-293 cells were cultivated in Dulbecco’s modified Eagle’s medium supplemented with 10% (vol/vol) fetal bovine serum, 4 mmol∙l-1 glutamine and 100 units∙ml-1 of both penicillin and streptomycin (Biochrom, Berlin, Germany).

    Techniques: Transferring, Blocking Assay

    Original recordings of two bTRPV3 cells filled with NMDG-gluconate solution. a) As before, replacement of chloride by gluconate resulted in a decrease in outward current at positive potentials, suggesting expression of chloride channels. Both addition of Ca 2+ (10 mmol∙l −1 ) and replacement by Ca-gluconate 2 (60 mmol∙l −1 ) had a strong blocking effect, with washout to levels higher than before. The current could be partially blocked by Mg 2+ , arguing against a rupture of the seal. Note the current kinetics with amplitude decreasing after a hyperpolarising pulse suggesting that Mg 2+ is being drawn into the channel pore by the negative membrane voltage. Conversely, a depolarising voltage pulse leads to a relief of the block. In both cases, the currents appear curved in contrast to the linear kinetics of currents in pure NMDG-gluconate solution. b) Second cell, showing a spontaneous activation of inward and outward current amplitude by a factor of about four between the first set of pulses in NMDG-gluconate EGTA solution (1) and a second set (2) that was run 100 seconds later. Current amplitude continued to increase both after addition of 10 mmol∙l −1 Ca 2+ to the NMDG-gluconate solution, and after replacement of NMDG-gluconate by Ca-gluconate 2 . Both the current kinetics and the tail currents indicate a partial voltage-dependent block. The arrows above the first set of pulses indicate the interval used for the determination of mean current amplitude for the current-voltage plot, which is depicted in c), showing rectification as a further sign of a voltage-dependent block. The reversal potential in 60 mmol∙l −1 Ca-gluconate 2 solution was 7.1 mV, revealing permeability to Ca 2+ .

    Journal: PLoS ONE

    Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+

    doi: 10.1371/journal.pone.0193519

    Figure Lengend Snippet: Original recordings of two bTRPV3 cells filled with NMDG-gluconate solution. a) As before, replacement of chloride by gluconate resulted in a decrease in outward current at positive potentials, suggesting expression of chloride channels. Both addition of Ca 2+ (10 mmol∙l −1 ) and replacement by Ca-gluconate 2 (60 mmol∙l −1 ) had a strong blocking effect, with washout to levels higher than before. The current could be partially blocked by Mg 2+ , arguing against a rupture of the seal. Note the current kinetics with amplitude decreasing after a hyperpolarising pulse suggesting that Mg 2+ is being drawn into the channel pore by the negative membrane voltage. Conversely, a depolarising voltage pulse leads to a relief of the block. In both cases, the currents appear curved in contrast to the linear kinetics of currents in pure NMDG-gluconate solution. b) Second cell, showing a spontaneous activation of inward and outward current amplitude by a factor of about four between the first set of pulses in NMDG-gluconate EGTA solution (1) and a second set (2) that was run 100 seconds later. Current amplitude continued to increase both after addition of 10 mmol∙l −1 Ca 2+ to the NMDG-gluconate solution, and after replacement of NMDG-gluconate by Ca-gluconate 2 . Both the current kinetics and the tail currents indicate a partial voltage-dependent block. The arrows above the first set of pulses indicate the interval used for the determination of mean current amplitude for the current-voltage plot, which is depicted in c), showing rectification as a further sign of a voltage-dependent block. The reversal potential in 60 mmol∙l −1 Ca-gluconate 2 solution was 7.1 mV, revealing permeability to Ca 2+ .

    Article Snippet: Cell culture and transient transfection of bTRPV3 HEK-293 cells were cultivated in Dulbecco’s modified Eagle’s medium supplemented with 10% (vol/vol) fetal bovine serum, 4 mmol∙l-1 glutamine and 100 units∙ml-1 of both penicillin and streptomycin (Biochrom, Berlin, Germany).

    Techniques: Expressing, Blocking Assay, Activation Assay, Permeability

    Response to various agonists of bTRPV3. Cells were filled with a CsCl pipette solution and stimulated with the continuous pulse protocol I. a) Original recording of a cell expressing bTRPV3, showing the response to application of menthol, thymol (both 1 mmol∙l −1 ), and 2-APB (0.3 mmol∙l −1 ) in NaCl bath solution. In response to 2-APB, the current rose dramatically, requiring an interruption of the measurement to readjust the gain (arrow). b) Original recording of a control cell transfected with the empty vector.

    Journal: PLoS ONE

    Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+

    doi: 10.1371/journal.pone.0193519

    Figure Lengend Snippet: Response to various agonists of bTRPV3. Cells were filled with a CsCl pipette solution and stimulated with the continuous pulse protocol I. a) Original recording of a cell expressing bTRPV3, showing the response to application of menthol, thymol (both 1 mmol∙l −1 ), and 2-APB (0.3 mmol∙l −1 ) in NaCl bath solution. In response to 2-APB, the current rose dramatically, requiring an interruption of the measurement to readjust the gain (arrow). b) Original recording of a control cell transfected with the empty vector.

    Article Snippet: Cell culture and transient transfection of bTRPV3 HEK-293 cells were cultivated in Dulbecco’s modified Eagle’s medium supplemented with 10% (vol/vol) fetal bovine serum, 4 mmol∙l-1 glutamine and 100 units∙ml-1 of both penicillin and streptomycin (Biochrom, Berlin, Germany).

    Techniques: Transferring, Expressing, Transfection, Plasmid Preparation

    Inside-out measurements from cells expressing bTRPV3 in symmetrical solutions. Panels (a to d) show recordings from the same patch and identical scaling in a) symmetrical NaCl solution, b) NaCl solution with 1.5 mmol∙l −1 MgCl 2 , c) Na-gluconate solution without MgCl 2 and d) return to symmetrical NaCl solution. The shift in the baseline at 0 mV pipette potential in (c) reflects the impact of the liquid junction potential. e) Patch from a second cell in symmetrical NH 4 Cl solution. For clarity, this graph only shows currents at every other voltage step of pulse protocol III. f) Current-voltage plot of the unitary current amplitudes of the data from (a to e) (linear fit).

    Journal: PLoS ONE

    Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+

    doi: 10.1371/journal.pone.0193519

    Figure Lengend Snippet: Inside-out measurements from cells expressing bTRPV3 in symmetrical solutions. Panels (a to d) show recordings from the same patch and identical scaling in a) symmetrical NaCl solution, b) NaCl solution with 1.5 mmol∙l −1 MgCl 2 , c) Na-gluconate solution without MgCl 2 and d) return to symmetrical NaCl solution. The shift in the baseline at 0 mV pipette potential in (c) reflects the impact of the liquid junction potential. e) Patch from a second cell in symmetrical NH 4 Cl solution. For clarity, this graph only shows currents at every other voltage step of pulse protocol III. f) Current-voltage plot of the unitary current amplitudes of the data from (a to e) (linear fit).

    Article Snippet: Cell culture and transient transfection of bTRPV3 HEK-293 cells were cultivated in Dulbecco’s modified Eagle’s medium supplemented with 10% (vol/vol) fetal bovine serum, 4 mmol∙l-1 glutamine and 100 units∙ml-1 of both penicillin and streptomycin (Biochrom, Berlin, Germany).

    Techniques: Expressing, Transferring

    Original recordings of two bTRPV3 cells filled with NMDG-gluconate solution. a) As before, replacement of chloride by gluconate resulted in a decrease in outward current at positive potentials, suggesting expression of chloride channels. Both addition of Ca 2+ (10 mmol∙l −1 ) and replacement by Ca-gluconate 2 (60 mmol∙l −1 ) had a strong blocking effect, with washout to levels higher than before. The current could be partially blocked by Mg 2+ , arguing against a rupture of the seal. Note the current kinetics with amplitude decreasing after a hyperpolarising pulse suggesting that Mg 2+ is being drawn into the channel pore by the negative membrane voltage. Conversely, a depolarising voltage pulse leads to a relief of the block. In both cases, the currents appear curved in contrast to the linear kinetics of currents in pure NMDG-gluconate solution. b) Second cell, showing a spontaneous activation of inward and outward current amplitude by a factor of about four between the first set of pulses in NMDG-gluconate EGTA solution (1) and a second set (2) that was run 100 seconds later. Current amplitude continued to increase both after addition of 10 mmol∙l −1 Ca 2+ to the NMDG-gluconate solution, and after replacement of NMDG-gluconate by Ca-gluconate 2 . Both the current kinetics and the tail currents indicate a partial voltage-dependent block. The arrows above the first set of pulses indicate the interval used for the determination of mean current amplitude for the current-voltage plot, which is depicted in c), showing rectification as a further sign of a voltage-dependent block. The reversal potential in 60 mmol∙l −1 Ca-gluconate 2 solution was 7.1 mV, revealing permeability to Ca 2+ .

    Journal: PLoS ONE

    Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+

    doi: 10.1371/journal.pone.0193519

    Figure Lengend Snippet: Original recordings of two bTRPV3 cells filled with NMDG-gluconate solution. a) As before, replacement of chloride by gluconate resulted in a decrease in outward current at positive potentials, suggesting expression of chloride channels. Both addition of Ca 2+ (10 mmol∙l −1 ) and replacement by Ca-gluconate 2 (60 mmol∙l −1 ) had a strong blocking effect, with washout to levels higher than before. The current could be partially blocked by Mg 2+ , arguing against a rupture of the seal. Note the current kinetics with amplitude decreasing after a hyperpolarising pulse suggesting that Mg 2+ is being drawn into the channel pore by the negative membrane voltage. Conversely, a depolarising voltage pulse leads to a relief of the block. In both cases, the currents appear curved in contrast to the linear kinetics of currents in pure NMDG-gluconate solution. b) Second cell, showing a spontaneous activation of inward and outward current amplitude by a factor of about four between the first set of pulses in NMDG-gluconate EGTA solution (1) and a second set (2) that was run 100 seconds later. Current amplitude continued to increase both after addition of 10 mmol∙l −1 Ca 2+ to the NMDG-gluconate solution, and after replacement of NMDG-gluconate by Ca-gluconate 2 . Both the current kinetics and the tail currents indicate a partial voltage-dependent block. The arrows above the first set of pulses indicate the interval used for the determination of mean current amplitude for the current-voltage plot, which is depicted in c), showing rectification as a further sign of a voltage-dependent block. The reversal potential in 60 mmol∙l −1 Ca-gluconate 2 solution was 7.1 mV, revealing permeability to Ca 2+ .

    Article Snippet: HEK-293 cells were cultivated in Dulbecco’s modified Eagle’s medium supplemented with 10% (vol/vol) fetal bovine serum, 4 mmol∙l-1 glutamine and 100 units∙ml-1 of both penicillin and streptomycin (Biochrom, Berlin, Germany).

    Techniques: Expressing, Blocking Assay, Activation Assay, Permeability

    Original recordings of one patch from a bTRPV3 cell with NH 4 Cl in the pipette. a) Channel events are clearly visible in NaCl bath solution in response to pulse protocol III both at negative potentials and positive potentials, reflecting influx of Na + and efflux of NH 4 + . b) same patch in 72.5 mmol∙l −1 CaCl 2 bath solution. Ca 2+ had a clear blocking effect on the current level at positive potentials (efflux of NH 4 + ). c) Detail from (b), showing single-channel events at negative potentials, reflecting influx of Ca 2+ d) same patch in NMDGCl bath solution. e) Insert showing single-channel events at negative potential level, suggesting influx of NMDG + . f) Current-voltage plot of unitary current amplitudes from this individual patch, fitted with the current formulation of the Goldman-Hodgkin-Katz equation. The fit yields a conductance for Na + of 132 ± 4 pS, for Ca 2+ of 21 ± 3 pS, and for NMDG + of 36 ± 3 pS. The conductance for NH 4 + was similar in NaCl and NMDGCl solution (250 ± 3 pS or 264 ± 3 pS, respectively) but dropped to 88 ± 4 pS with high Ca 2+ in the bath.

    Journal: PLoS ONE

    Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+

    doi: 10.1371/journal.pone.0193519

    Figure Lengend Snippet: Original recordings of one patch from a bTRPV3 cell with NH 4 Cl in the pipette. a) Channel events are clearly visible in NaCl bath solution in response to pulse protocol III both at negative potentials and positive potentials, reflecting influx of Na + and efflux of NH 4 + . b) same patch in 72.5 mmol∙l −1 CaCl 2 bath solution. Ca 2+ had a clear blocking effect on the current level at positive potentials (efflux of NH 4 + ). c) Detail from (b), showing single-channel events at negative potentials, reflecting influx of Ca 2+ d) same patch in NMDGCl bath solution. e) Insert showing single-channel events at negative potential level, suggesting influx of NMDG + . f) Current-voltage plot of unitary current amplitudes from this individual patch, fitted with the current formulation of the Goldman-Hodgkin-Katz equation. The fit yields a conductance for Na + of 132 ± 4 pS, for Ca 2+ of 21 ± 3 pS, and for NMDG + of 36 ± 3 pS. The conductance for NH 4 + was similar in NaCl and NMDGCl solution (250 ± 3 pS or 264 ± 3 pS, respectively) but dropped to 88 ± 4 pS with high Ca 2+ in the bath.

    Article Snippet: HEK-293 cells were cultivated in Dulbecco’s modified Eagle’s medium supplemented with 10% (vol/vol) fetal bovine serum, 4 mmol∙l-1 glutamine and 100 units∙ml-1 of both penicillin and streptomycin (Biochrom, Berlin, Germany).

    Techniques: Transferring, Blocking Assay

    Effects of menthol and Mg 2+ on intracellular calcium [Ca 2+ ] i . a) Average of ten original recordings of control HEK-293 cells, showing a significant increase of Δ [Ca 2+ ] i in the 200 s interval after application of menthol (1 mmol∙l −1 ) (95% confidence interval in grey), with the timeline identical to that in b), showing the average [Ca 2+ ] i (± 95%) of eleven recordings of HEK-293 cells overexpressing bTRPV3. While resting levels of [Ca 2+ ] i in bTRPV3 cells were not significantly different from those of control cells, menthol led to a significantly higher Δ [Ca 2+ ] i in these cells (p = 0.003). c) Boxplots comparing the slopes (Δ [Ca 2+ ] i /Δ t) of the graphs in (a and b) in a 50 s interval at the beginning of the measurement and in the 50 s interval after application of menthol (differing letters above the boxes indicate p

    Journal: PLoS ONE

    Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+

    doi: 10.1371/journal.pone.0193519

    Figure Lengend Snippet: Effects of menthol and Mg 2+ on intracellular calcium [Ca 2+ ] i . a) Average of ten original recordings of control HEK-293 cells, showing a significant increase of Δ [Ca 2+ ] i in the 200 s interval after application of menthol (1 mmol∙l −1 ) (95% confidence interval in grey), with the timeline identical to that in b), showing the average [Ca 2+ ] i (± 95%) of eleven recordings of HEK-293 cells overexpressing bTRPV3. While resting levels of [Ca 2+ ] i in bTRPV3 cells were not significantly different from those of control cells, menthol led to a significantly higher Δ [Ca 2+ ] i in these cells (p = 0.003). c) Boxplots comparing the slopes (Δ [Ca 2+ ] i /Δ t) of the graphs in (a and b) in a 50 s interval at the beginning of the measurement and in the 50 s interval after application of menthol (differing letters above the boxes indicate p

    Article Snippet: HEK-293 cells were cultivated in Dulbecco’s modified Eagle’s medium supplemented with 10% (vol/vol) fetal bovine serum, 4 mmol∙l-1 glutamine and 100 units∙ml-1 of both penicillin and streptomycin (Biochrom, Berlin, Germany).

    Techniques:

    Inside-out measurements from cells expressing bTRPV3 in symmetrical solutions. Panels (a to d) show recordings from the same patch and identical scaling in a) symmetrical NaCl solution, b) NaCl solution with 1.5 mmol∙l −1 MgCl 2 , c) Na-gluconate solution without MgCl 2 and d) return to symmetrical NaCl solution. The shift in the baseline at 0 mV pipette potential in (c) reflects the impact of the liquid junction potential. e) Patch from a second cell in symmetrical NH 4 Cl solution. For clarity, this graph only shows currents at every other voltage step of pulse protocol III. f) Current-voltage plot of the unitary current amplitudes of the data from (a to e) (linear fit).

    Journal: PLoS ONE

    Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+

    doi: 10.1371/journal.pone.0193519

    Figure Lengend Snippet: Inside-out measurements from cells expressing bTRPV3 in symmetrical solutions. Panels (a to d) show recordings from the same patch and identical scaling in a) symmetrical NaCl solution, b) NaCl solution with 1.5 mmol∙l −1 MgCl 2 , c) Na-gluconate solution without MgCl 2 and d) return to symmetrical NaCl solution. The shift in the baseline at 0 mV pipette potential in (c) reflects the impact of the liquid junction potential. e) Patch from a second cell in symmetrical NH 4 Cl solution. For clarity, this graph only shows currents at every other voltage step of pulse protocol III. f) Current-voltage plot of the unitary current amplitudes of the data from (a to e) (linear fit).

    Article Snippet: HEK-293 cells were cultivated in Dulbecco’s modified Eagle’s medium supplemented with 10% (vol/vol) fetal bovine serum, 4 mmol∙l-1 glutamine and 100 units∙ml-1 of both penicillin and streptomycin (Biochrom, Berlin, Germany).

    Techniques: Expressing, Transferring