hepes buffer  (Tocris)

 
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  • 99
    Name:
    HEPES
    Description:
    Multipurpose buffer
    Catalog Number:
    3173
    Price:
    None
    Purity:
    ≥99.5%
    Category:
    Buffers Solvents and Solutions Reagents
    Formula:
    4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid
    Buy from Supplier


    Structured Review

    Tocris hepes buffer
    HEPES
    Multipurpose buffer
    https://www.bioz.com/result/hepes buffer/product/Tocris
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hepes buffer - by Bioz Stars, 2020-04
    99/100 stars

    Images

    1) Product Images from "Glutamate Uptake Triggers Transporter-Mediated GABA Release from Astrocytes"

    Article Title: Glutamate Uptake Triggers Transporter-Mediated GABA Release from Astrocytes

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0007153

    Glu-induced GABA release is not mediated by vesicular release. (A) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in the presence of glutamate receptor antagonists (n = 7; P = 0.014; 128.5±13.4% of pre-stimulus control). (B) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 and Glu receptor antagonists in rat cerebrocortical NPMV fractions (n = 4; P = 0.82) (C) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in low-[Ca 2+ ]/high-[Mg 2+ ] buffer (n = 5; P = 0.91; 148.3±15.5% of pre-stimulus control). (D) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 in low-[Ca 2+ ] buffer in rat cerebrocortical NPMV fractions (n = 3; P = 0.49). Low-[Ca 2+ ] buffer contained 145 mM NaCl, 5 mM KCl, 20 mM MgCl 2 , 10 mM glucose and 20 mM HEPES (pH 7.5).
    Figure Legend Snippet: Glu-induced GABA release is not mediated by vesicular release. (A) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in the presence of glutamate receptor antagonists (n = 7; P = 0.014; 128.5±13.4% of pre-stimulus control). (B) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 and Glu receptor antagonists in rat cerebrocortical NPMV fractions (n = 4; P = 0.82) (C) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in low-[Ca 2+ ]/high-[Mg 2+ ] buffer (n = 5; P = 0.91; 148.3±15.5% of pre-stimulus control). (D) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 in low-[Ca 2+ ] buffer in rat cerebrocortical NPMV fractions (n = 3; P = 0.49). Low-[Ca 2+ ] buffer contained 145 mM NaCl, 5 mM KCl, 20 mM MgCl 2 , 10 mM glucose and 20 mM HEPES (pH 7.5).

    Techniques Used:

    Related Articles

    Conditioned Place Preference:

    Article Title: NPY Y1 Receptors Differentially Modulate GABAA and NMDA Receptors via Divergent Signal-Transduction Pathways to Reduce Excitability of Amygdala Neurons
    Article Snippet: .. The following drugs were obtained from (1) Sigma-Aldrich (St Louis, MO): K-gluconate, KCl, MgCl2 , HEPES, NaCl, KH2 PO4 , CaCl2 , glucose, NaHCO3 , KOH, phosphocreatine, K-ATP, Na-GTP, DNQX, 8Br-cAMP, GDP- β -S; (2) Tocris Cookson (Ellisville, MO): [Leu31 -Pro34 ]-NPY, PD160170, ( RS )-CPP, CGP 52432, SR95531, 8CPT-2Me-cAMP, forskolin, QX-314; and (3) EMD Chemicals (Gibbstown, NJ): protein kinase A (PKA) catalytic subunit. .. BLA, hippocampus, and PFC tissue punches from rats, or CHOK1 and HEK293 cells, were lysed with buffer (30 mM Tris, pH 7.4, 150 mM NaCl, 1% Triton X-100, 0.1% SDS, 1 mM PMSF, 10 mM EDTA, 1 mM Na2 VO3 , and 160 mM NaF) freshly supplemented with proteinase inhibitors, and protein concentrations were determined using a BCA Protein Assay Kit (Thermo Fisher Scientific, Rockford, IL).

    Flow Cytometry:


    Permeability:

    Article Title: UCP2 modulates single-channel properties of a MCU-dependent Ca2+ inward current in mitochondria
    Article Snippet: .. For single-channel recordings, the pipette solution contained 105 mM CaCl2 and 10 mM HEPES, 10 μM cyclosporin A (Tocris Bioscience, Bristol, UK) and 10 μM 7-chloro-5-(2-chlorophenyl)-1,5-dihydro-4,1-benzothiazepin-2(3H )-one (CGP 37157, Ascent Scientific Ltd., Bristol, UK) to prevent opening of the permeability transition pore (PTP), and the activity of the mitochondrial Na+ /Ca2+ exchanger (NCXmito ), respectively. pH was adjusted to 7.2 with Ca(OH)2 . ..

    Article Title: Mitochondrial Ca2+ uniporter (MCU)-dependent and MCU-independent Ca2+ channels coexist in the inner mitochondrial membrane
    Article Snippet: .. For single channel recordings, the pipette solution contained 105 mM CaCl2 and 10 mM HEPES, 10 μM cyclosporin A (Tocris Bioscience, Bristol, UK) and 10 μM 7-chloro-5-(2-chlorophenyl)-1,5-dihydro-4,1-benzothiazepin-2(3H )-one (CGP 37157, Ascent Scientific Ltd., Bristol, UK) to prevent opening of the permeability transition pore, and the activity of the mitochondrial Na+ /Ca2+ exchanger (NCXmito ), respectively. pH was adjusted to 7.2 with Ca(OH)2 . ..

    Patch Clamp:

    Article Title: UCP2 modulates single-channel properties of a MCU-dependent Ca2+ inward current in mitochondria
    Article Snippet: Paragraph title: Mitoplast patch clamp recordings ... For single-channel recordings, the pipette solution contained 105 mM CaCl2 and 10 mM HEPES, 10 μM cyclosporin A (Tocris Bioscience, Bristol, UK) and 10 μM 7-chloro-5-(2-chlorophenyl)-1,5-dihydro-4,1-benzothiazepin-2(3H )-one (CGP 37157, Ascent Scientific Ltd., Bristol, UK) to prevent opening of the permeability transition pore (PTP), and the activity of the mitochondrial Na+ /Ca2+ exchanger (NCXmito ), respectively. pH was adjusted to 7.2 with Ca(OH)2 .

    Article Title: Functional analysis of the interface between the tandem C2 domains of synaptotagmin-1
    Article Snippet: Electrophysiology Whole-cell patch-clamp recordings of mEPSCs, evoked EPSCs, and hypertonic sucrose responses from neurons at 14–15 DIV, were performed in voltage-clamp mode using an EPC-10/2 amplifier (HEKA, Holliston, MA). .. The bath solution consisted of 128 mM NaCl, 30 mM glucose, 5 mM KCl, 5 mM CaCl2 , 1 mM MgCl2 , 50 mM d -AP5, 20 mM bicuculline, and 25 mM HEPES (pH 7.3). d -AP5, bicuculline, and QX-314 were obtained from TOCRIS Bioscience (Bristol, United Kingdom).

    Article Title: Soluble TREM2 ameliorates pathological phenotypes by modulating microglial functions in an Alzheimer’s disease model
    Article Snippet: The patch pipette (5–8 MΩ) was filled with a standard intracellular solution containing (in mM) 140 CsCH3 SO3 , 2 MgCl·6H2 O, 5 TEA-Cl, 10 HEPES, 2.5 MgATP, 0.3 Na2 GTP, and 1 EGTA, pH 7.2–7.4 (with CsOH). mEPSCs and mIPSCs were recorded with 1 μM tetrodotoxin (TTX; Tocris, Cat. No. 1069) in voltage-clamp mode at a holding potential of −70 mV and 0 mV, respectively. .. Whole-cell recording was made using patch-clamp amplifiers (Multiclamp 700B).

    Article Title: Maternal Care Differentially Affects Neuronal Excitability and Synaptic Plasticity in the Dorsal and Ventral Hippocampus
    Article Snippet: .. To record α -Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor-mediated miniature excitatory postsynaptic currents (mEPSCs), whole-cell patch clamp recording of CA1 pyramidal neurons was performed in voltage clamp mode with pipettes containing (in mM): 110 Cs-gluconate, 17.5 CsCl, 10 HEPES, 2 MgCl2 , 0.5 EGTA, 4 Na2 -ATP, 5 QX-314, and 0.5% Biocytin (Tocris) with the pH adjusted to 7.2 with CsOH (∼290 mOsm). .. Neurons were held at −60 mV, and recordings of mEPSCs were performed in the presence of tetrodotoxin (0.5 μM; Alomone Labs, Jerusalem, Israel) to block action-potential formation, and 5 μM bicuculline (Tocris) to block GABAA receptor-mediated inhibitory currents.

    Article Title: Mitochondrial Ca2+ uniporter (MCU)-dependent and MCU-independent Ca2+ channels coexist in the inner mitochondrial membrane
    Article Snippet: Paragraph title: Mitoplast patch-clamp recordings ... For single channel recordings, the pipette solution contained 105 mM CaCl2 and 10 mM HEPES, 10 μM cyclosporin A (Tocris Bioscience, Bristol, UK) and 10 μM 7-chloro-5-(2-chlorophenyl)-1,5-dihydro-4,1-benzothiazepin-2(3H )-one (CGP 37157, Ascent Scientific Ltd., Bristol, UK) to prevent opening of the permeability transition pore, and the activity of the mitochondrial Na+ /Ca2+ exchanger (NCXmito ), respectively. pH was adjusted to 7.2 with Ca(OH)2 .

    Transferring:

    Article Title: UCP2 modulates single-channel properties of a MCU-dependent Ca2+ inward current in mitochondria
    Article Snippet: .. For single-channel recordings, the pipette solution contained 105 mM CaCl2 and 10 mM HEPES, 10 μM cyclosporin A (Tocris Bioscience, Bristol, UK) and 10 μM 7-chloro-5-(2-chlorophenyl)-1,5-dihydro-4,1-benzothiazepin-2(3H )-one (CGP 37157, Ascent Scientific Ltd., Bristol, UK) to prevent opening of the permeability transition pore (PTP), and the activity of the mitochondrial Na+ /Ca2+ exchanger (NCXmito ), respectively. pH was adjusted to 7.2 with Ca(OH)2 . ..

    Article Title: Auxiliary Subunit GSG1L Acts to Suppress Calcium-Permeable AMPA Receptor Function
    Article Snippet: .. The internal (pipette) solution contained the following (in m m ): 145 CsCl, 2.5 NaCl, 1 Cs-EGTA, 4 Mg-ATP, 10 HEPES, pH 7.3 with CsOH, and 0.1 spermine tetrahydrochloride (Tocris Bioscience). ..

    Article Title: Functional analysis of the interface between the tandem C2 domains of synaptotagmin-1
    Article Snippet: The pipette solution consisted of 130 mM K-gluconate, 1 mM EGTA, 5 mM Na-phosphocreatine, 2 mM Mg-ATP, 0.3 mM Na-GTP, 5 mM QX-314, and 10 mM HEPES (pH 7.3). .. The bath solution consisted of 128 mM NaCl, 30 mM glucose, 5 mM KCl, 5 mM CaCl2 , 1 mM MgCl2 , 50 mM d -AP5, 20 mM bicuculline, and 25 mM HEPES (pH 7.3). d -AP5, bicuculline, and QX-314 were obtained from TOCRIS Bioscience (Bristol, United Kingdom).

    Article Title: Soluble TREM2 ameliorates pathological phenotypes by modulating microglial functions in an Alzheimer’s disease model
    Article Snippet: .. The patch pipette (5–8 MΩ) was filled with a standard intracellular solution containing (in mM) 140 CsCH3 SO3 , 2 MgCl·6H2 O, 5 TEA-Cl, 10 HEPES, 2.5 MgATP, 0.3 Na2 GTP, and 1 EGTA, pH 7.2–7.4 (with CsOH). mEPSCs and mIPSCs were recorded with 1 μM tetrodotoxin (TTX; Tocris, Cat. No. 1069) in voltage-clamp mode at a holding potential of −70 mV and 0 mV, respectively. ..

    Article Title: Mitochondrial Ca2+ uniporter (MCU)-dependent and MCU-independent Ca2+ channels coexist in the inner mitochondrial membrane
    Article Snippet: .. For single channel recordings, the pipette solution contained 105 mM CaCl2 and 10 mM HEPES, 10 μM cyclosporin A (Tocris Bioscience, Bristol, UK) and 10 μM 7-chloro-5-(2-chlorophenyl)-1,5-dihydro-4,1-benzothiazepin-2(3H )-one (CGP 37157, Ascent Scientific Ltd., Bristol, UK) to prevent opening of the permeability transition pore, and the activity of the mitochondrial Na+ /Ca2+ exchanger (NCXmito ), respectively. pH was adjusted to 7.2 with Ca(OH)2 . ..

    Cytometry:

    Article Title: P2X7 Receptors Regulate Phagocytosis and Proliferation in Adult Hippocampal and SVZ Neural Progenitor Cells: Implications for Inflammation in Neurogenesis
    Article Snippet: NPCs were washed twice with Ca2+ ‐free Na+ buffer and allowed to de‐esterify on ice for 30 minutes before resuspension in K+ buffer (in mM: KCl [145], KOH [5], HEPES [10], glucose [5], CaCl2 [0.1], BSA [1%]) with or without preincubation with the P2X7 specific inhibitors AZ10606120 (15 minutes, 1 μM, Tocris Bioscience) or A438079 (30 minutes, 10 μM, Tocris Biosciences, Bristol). .. CaCl2 (3 mM) was added prior to analysis by flow cytometry, followed by the addition of ATP (1 mM) or BzATP (100 μM) after 40 seconds of acquisition.

    Blocking Assay:

    Article Title: Cumulative induction of glycinergic inhibitory plasticity in binaural neurons is gated by developmental changes in action potential backpropagation
    Article Snippet: .. For the voltage-clamp experiments testing glutamate co-release and GluN2B subunit block, the internal solution contained (in mM): 45 CsMeSO3 , 80 CsCl, 0.5, 1.8 CaCl2 , 5 EGTA, 10 HEPES, 10 Na2 phosphocreatine, ~14 sucrose, 2 Mg-ATP, 0.3 Na-GTP, 1.46 QX-314 (Br, Tocris), 0.02 ZD 7288 (Tocris), pH 7.30 with CsOH, ~305 mmol/kg. ..

    Article Title: Lack of Synapsin I Reduces the Readily Releasable Pool of Synaptic Vesicles at Central Inhibitory Synapses
    Article Snippet: .. mIPSCs and eIPSCs were recorded by superfusing the whole-cell clamped postsynaptic neuron with a Tyrode solution containing (in m m ) 2 CaCl2 , 150 NaCl, 1 MgCl2 , 10 HEPES, 4 KCl, 10 glucose, pH 7.4. d -(−)-2-amino-5-phosphonopentanoic acid ( d -AP5; 50 μ m ), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 10 μ m ), and (5 μ m ) (Tocris, Bristol, UK) were added to the Tyrode solution to block NMDA, non-NMDA, and GABAB receptors, respectively. .. In some cases, the nonselective glutamate receptor antagonist kynurenic acid (1 m m ; Sigma, St. Louis, MO) was used to block excitatory transmission.

    Article Title: Maternal Care Differentially Affects Neuronal Excitability and Synaptic Plasticity in the Dorsal and Ventral Hippocampus
    Article Snippet: Field recordings were performed in the presence of 5 μM bicuculline methobromide (Tocris, Bristol, UK) and 5 μM picrotoxin (Tocris) to block γ -Aminobutyric acid type A (GABAA ) receptor-mediated inhibitory postsynaptic transmission unless otherwise specified. .. To record α -Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor-mediated miniature excitatory postsynaptic currents (mEPSCs), whole-cell patch clamp recording of CA1 pyramidal neurons was performed in voltage clamp mode with pipettes containing (in mM): 110 Cs-gluconate, 17.5 CsCl, 10 HEPES, 2 MgCl2 , 0.5 EGTA, 4 Na2 -ATP, 5 QX-314, and 0.5% Biocytin (Tocris) with the pH adjusted to 7.2 with CsOH (∼290 mOsm).

    Slice Preparation:

    Article Title: NPY Y1 Receptors Differentially Modulate GABAA and NMDA Receptors via Divergent Signal-Transduction Pathways to Reduce Excitability of Amygdala Neurons
    Article Snippet: Various drugs, diluted to the final concentration in ACSF, were delivered by gravity to the recording chamber containing the brain slice using a VC-6 control perfusion system (Harvard Apparatus, Holliston, MA), unless stated otherwise. .. The following drugs were obtained from (1) Sigma-Aldrich (St Louis, MO): K-gluconate, KCl, MgCl2 , HEPES, NaCl, KH2 PO4 , CaCl2 , glucose, NaHCO3 , KOH, phosphocreatine, K-ATP, Na-GTP, DNQX, 8Br-cAMP, GDP- β -S; (2) Tocris Cookson (Ellisville, MO): [Leu31 -Pro34 ]-NPY, PD160170, ( RS )-CPP, CGP 52432, SR95531, 8CPT-2Me-cAMP, forskolin, QX-314; and (3) EMD Chemicals (Gibbstown, NJ): protein kinase A (PKA) catalytic subunit.

    Concentration Assay:

    Article Title: NPY Y1 Receptors Differentially Modulate GABAA and NMDA Receptors via Divergent Signal-Transduction Pathways to Reduce Excitability of Amygdala Neurons
    Article Snippet: Various drugs, diluted to the final concentration in ACSF, were delivered by gravity to the recording chamber containing the brain slice using a VC-6 control perfusion system (Harvard Apparatus, Holliston, MA), unless stated otherwise. .. The following drugs were obtained from (1) Sigma-Aldrich (St Louis, MO): K-gluconate, KCl, MgCl2 , HEPES, NaCl, KH2 PO4 , CaCl2 , glucose, NaHCO3 , KOH, phosphocreatine, K-ATP, Na-GTP, DNQX, 8Br-cAMP, GDP- β -S; (2) Tocris Cookson (Ellisville, MO): [Leu31 -Pro34 ]-NPY, PD160170, ( RS )-CPP, CGP 52432, SR95531, 8CPT-2Me-cAMP, forskolin, QX-314; and (3) EMD Chemicals (Gibbstown, NJ): protein kinase A (PKA) catalytic subunit.

    Incubation:

    Article Title: Phosphorylation of IRS1 at Serine 307 in Response to Insulin in Human Adipocytes Is Not Likely to be Catalyzed by p70 Ribosomal S6 Kinase
    Article Snippet: .. Ser307-kinase Assay Immunoprecipitates of mTOR were incubated with the S307-peptide and 50 µM ATP in the presence of 10 mM MgCl2 , 50 mM KCl and 25 mM Hepes at pH 7.4 for 2 h at 30°C, with or without 0.5 µM Torin (Tocris Bioscience, Bristol, UK), 20 µM PF-4708671 (PZ0143, Sigma Chem. .. Co), or 3 µM Akti1/2 (sc-202048 Santa Cruz Biotechnology, Santa Cruz, CA, USA), as indicated.

    Article Title: Glutamate Uptake Triggers Transporter-Mediated GABA Release from Astrocytes
    Article Snippet: Uptake studies were performed as follows: aliquots of NPMV fractions in HEPES buffer were preincubated with test compounds in the presence of NNC-711 (100 µM, Tocris) at 30°C for 10 min. .. Afterward, preincubated solution was incubated with [3 H]GABA (10 nM, Amersham) and GABA (90 nM, Sigma) for 1 min.

    Article Title: Soluble TREM2 ameliorates pathological phenotypes by modulating microglial functions in an Alzheimer’s disease model
    Article Snippet: For sTREM2 protein treatment, neurons were incubated with 50 nM recombinant sTREM2-Fc or Fc protein prepared as previously described for 1 h, and transferred to the chamber for analysis. .. The patch pipette (5–8 MΩ) was filled with a standard intracellular solution containing (in mM) 140 CsCH3 SO3 , 2 MgCl·6H2 O, 5 TEA-Cl, 10 HEPES, 2.5 MgATP, 0.3 Na2 GTP, and 1 EGTA, pH 7.2–7.4 (with CsOH). mEPSCs and mIPSCs were recorded with 1 μM tetrodotoxin (TTX; Tocris, Cat. No. 1069) in voltage-clamp mode at a holding potential of −70 mV and 0 mV, respectively.

    Activity Assay:

    Article Title: UCP2 modulates single-channel properties of a MCU-dependent Ca2+ inward current in mitochondria
    Article Snippet: .. For single-channel recordings, the pipette solution contained 105 mM CaCl2 and 10 mM HEPES, 10 μM cyclosporin A (Tocris Bioscience, Bristol, UK) and 10 μM 7-chloro-5-(2-chlorophenyl)-1,5-dihydro-4,1-benzothiazepin-2(3H )-one (CGP 37157, Ascent Scientific Ltd., Bristol, UK) to prevent opening of the permeability transition pore (PTP), and the activity of the mitochondrial Na+ /Ca2+ exchanger (NCXmito ), respectively. pH was adjusted to 7.2 with Ca(OH)2 . ..

    Article Title: Mitochondrial Ca2+ uniporter (MCU)-dependent and MCU-independent Ca2+ channels coexist in the inner mitochondrial membrane
    Article Snippet: .. For single channel recordings, the pipette solution contained 105 mM CaCl2 and 10 mM HEPES, 10 μM cyclosporin A (Tocris Bioscience, Bristol, UK) and 10 μM 7-chloro-5-(2-chlorophenyl)-1,5-dihydro-4,1-benzothiazepin-2(3H )-one (CGP 37157, Ascent Scientific Ltd., Bristol, UK) to prevent opening of the permeability transition pore, and the activity of the mitochondrial Na+ /Ca2+ exchanger (NCXmito ), respectively. pH was adjusted to 7.2 with Ca(OH)2 . ..

    Cell Culture:

    Article Title: Soluble TREM2 ameliorates pathological phenotypes by modulating microglial functions in an Alzheimer’s disease model
    Article Snippet: Whole-cell voltage-clamp recordings were performed on cortical neurons cultured for 12–14 DIV . .. The patch pipette (5–8 MΩ) was filled with a standard intracellular solution containing (in mM) 140 CsCH3 SO3 , 2 MgCl·6H2 O, 5 TEA-Cl, 10 HEPES, 2.5 MgATP, 0.3 Na2 GTP, and 1 EGTA, pH 7.2–7.4 (with CsOH). mEPSCs and mIPSCs were recorded with 1 μM tetrodotoxin (TTX; Tocris, Cat. No. 1069) in voltage-clamp mode at a holding potential of −70 mV and 0 mV, respectively.

    Mass Spectrometry:

    Article Title: Maternal Care Differentially Affects Neuronal Excitability and Synaptic Plasticity in the Dorsal and Ventral Hippocampus
    Article Snippet: Neurons recorded in current clamp mode were injected with 1000 ms long depolarizing current pulses in increasing steps of 10 pA to evoke action potentials. .. To record α -Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor-mediated miniature excitatory postsynaptic currents (mEPSCs), whole-cell patch clamp recording of CA1 pyramidal neurons was performed in voltage clamp mode with pipettes containing (in mM): 110 Cs-gluconate, 17.5 CsCl, 10 HEPES, 2 MgCl2 , 0.5 EGTA, 4 Na2 -ATP, 5 QX-314, and 0.5% Biocytin (Tocris) with the pH adjusted to 7.2 with CsOH (∼290 mOsm).

    Article Title: Mitochondrial Ca2+ uniporter (MCU)-dependent and MCU-independent Ca2+ channels coexist in the inner mitochondrial membrane
    Article Snippet: For single channel recordings, the pipette solution contained 105 mM CaCl2 and 10 mM HEPES, 10 μM cyclosporin A (Tocris Bioscience, Bristol, UK) and 10 μM 7-chloro-5-(2-chlorophenyl)-1,5-dihydro-4,1-benzothiazepin-2(3H )-one (CGP 37157, Ascent Scientific Ltd., Bristol, UK) to prevent opening of the permeability transition pore, and the activity of the mitochondrial Na+ /Ca2+ exchanger (NCXmito ), respectively. pH was adjusted to 7.2 with Ca(OH)2 . .. For obtaining whole-mitoplast configuration, voltage steps of 300–600 mV and 20–50 ms duration were applied.

    Transmission Assay:

    Article Title: Lack of Synapsin I Reduces the Readily Releasable Pool of Synaptic Vesicles at Central Inhibitory Synapses
    Article Snippet: mIPSCs and eIPSCs were recorded by superfusing the whole-cell clamped postsynaptic neuron with a Tyrode solution containing (in m m ) 2 CaCl2 , 150 NaCl, 1 MgCl2 , 10 HEPES, 4 KCl, 10 glucose, pH 7.4. d -(−)-2-amino-5-phosphonopentanoic acid ( d -AP5; 50 μ m ), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 10 μ m ), and (5 μ m ) (Tocris, Bristol, UK) were added to the Tyrode solution to block NMDA, non-NMDA, and GABAB receptors, respectively. .. In some cases, the nonselective glutamate receptor antagonist kynurenic acid (1 m m ; Sigma, St. Louis, MO) was used to block excitatory transmission.

    Article Title: Soluble TREM2 ameliorates pathological phenotypes by modulating microglial functions in an Alzheimer’s disease model
    Article Snippet: The strength of synaptic transmission was determined by measuring the initial (20–80% rising phase) slope of fEPSPs. .. The patch pipette (5–8 MΩ) was filled with a standard intracellular solution containing (in mM) 140 CsCH3 SO3 , 2 MgCl·6H2 O, 5 TEA-Cl, 10 HEPES, 2.5 MgATP, 0.3 Na2 GTP, and 1 EGTA, pH 7.2–7.4 (with CsOH). mEPSCs and mIPSCs were recorded with 1 μM tetrodotoxin (TTX; Tocris, Cat. No. 1069) in voltage-clamp mode at a holding potential of −70 mV and 0 mV, respectively.

    Article Title: Maternal Care Differentially Affects Neuronal Excitability and Synaptic Plasticity in the Dorsal and Ventral Hippocampus
    Article Snippet: Field recordings were performed in the presence of 5 μM bicuculline methobromide (Tocris, Bristol, UK) and 5 μM picrotoxin (Tocris) to block γ -Aminobutyric acid type A (GABAA ) receptor-mediated inhibitory postsynaptic transmission unless otherwise specified. .. To record α -Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor-mediated miniature excitatory postsynaptic currents (mEPSCs), whole-cell patch clamp recording of CA1 pyramidal neurons was performed in voltage clamp mode with pipettes containing (in mM): 110 Cs-gluconate, 17.5 CsCl, 10 HEPES, 2 MgCl2 , 0.5 EGTA, 4 Na2 -ATP, 5 QX-314, and 0.5% Biocytin (Tocris) with the pH adjusted to 7.2 with CsOH (∼290 mOsm).

    Injection:

    Article Title: Maternal Care Differentially Affects Neuronal Excitability and Synaptic Plasticity in the Dorsal and Ventral Hippocampus
    Article Snippet: Neurons recorded in current clamp mode were injected with 1000 ms long depolarizing current pulses in increasing steps of 10 pA to evoke action potentials. .. To record α -Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor-mediated miniature excitatory postsynaptic currents (mEPSCs), whole-cell patch clamp recording of CA1 pyramidal neurons was performed in voltage clamp mode with pipettes containing (in mM): 110 Cs-gluconate, 17.5 CsCl, 10 HEPES, 2 MgCl2 , 0.5 EGTA, 4 Na2 -ATP, 5 QX-314, and 0.5% Biocytin (Tocris) with the pH adjusted to 7.2 with CsOH (∼290 mOsm).

    Recombinant:

    Article Title: Soluble TREM2 ameliorates pathological phenotypes by modulating microglial functions in an Alzheimer’s disease model
    Article Snippet: For sTREM2 protein treatment, neurons were incubated with 50 nM recombinant sTREM2-Fc or Fc protein prepared as previously described for 1 h, and transferred to the chamber for analysis. .. The patch pipette (5–8 MΩ) was filled with a standard intracellular solution containing (in mM) 140 CsCH3 SO3 , 2 MgCl·6H2 O, 5 TEA-Cl, 10 HEPES, 2.5 MgATP, 0.3 Na2 GTP, and 1 EGTA, pH 7.2–7.4 (with CsOH). mEPSCs and mIPSCs were recorded with 1 μM tetrodotoxin (TTX; Tocris, Cat. No. 1069) in voltage-clamp mode at a holding potential of −70 mV and 0 mV, respectively.

    Software:

    Article Title: Auxiliary Subunit GSG1L Acts to Suppress Calcium-Permeable AMPA Receptor Function
    Article Snippet: Records were low-pass filtered at 5 kHz and digitized at 20 kHz using WINWCP software (John Dempster, University of Strathclyde, Glasgow, UK) and analyzed using Igor Pro 6.35 (WaveMetrics) with NeuroMatic 2.8v ( ). .. The internal (pipette) solution contained the following (in m m ): 145 CsCl, 2.5 NaCl, 1 Cs-EGTA, 4 Mg-ATP, 10 HEPES, pH 7.3 with CsOH, and 0.1 spermine tetrahydrochloride (Tocris Bioscience).

    Article Title: Soluble TREM2 ameliorates pathological phenotypes by modulating microglial functions in an Alzheimer’s disease model
    Article Snippet: The patch pipette (5–8 MΩ) was filled with a standard intracellular solution containing (in mM) 140 CsCH3 SO3 , 2 MgCl·6H2 O, 5 TEA-Cl, 10 HEPES, 2.5 MgATP, 0.3 Na2 GTP, and 1 EGTA, pH 7.2–7.4 (with CsOH). mEPSCs and mIPSCs were recorded with 1 μM tetrodotoxin (TTX; Tocris, Cat. No. 1069) in voltage-clamp mode at a holding potential of −70 mV and 0 mV, respectively. .. Data acquisition was performed using digitizers (DigiData 1440A). mEPSC and mIPSC were analyzed off-line with MiniAnalysis software (Synaptosoft Inc., Fort Lee, NJ).

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  • 99
    Tocris hepes buffer
    Glu-induced GABA release is not mediated by vesicular release. (A) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in the presence of glutamate receptor antagonists (n = 7; P = 0.014; 128.5±13.4% of pre-stimulus control). (B) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 and Glu receptor antagonists in rat cerebrocortical <t>NPMV</t> fractions (n = 4; P = 0.82) (C) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in low-[Ca 2+ ]/high-[Mg 2+ ] buffer (n = 5; P = 0.91; 148.3±15.5% of pre-stimulus control). (D) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 in low-[Ca 2+ ] buffer in rat cerebrocortical NPMV fractions (n = 3; P = 0.49). Low-[Ca 2+ ] buffer contained 145 mM NaCl, 5 mM KCl, 20 mM MgCl 2 , 10 mM glucose and 20 mM <t>HEPES</t> (pH 7.5).
    Hepes Buffer, supplied by Tocris, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Tocris hepes buffered acsf
    Glu-induced GABA release is not mediated by vesicular release. (A) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in the presence of glutamate receptor antagonists (n = 7; P = 0.014; 128.5±13.4% of pre-stimulus control). (B) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 and Glu receptor antagonists in rat cerebrocortical <t>NPMV</t> fractions (n = 4; P = 0.82) (C) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in low-[Ca 2+ ]/high-[Mg 2+ ] buffer (n = 5; P = 0.91; 148.3±15.5% of pre-stimulus control). (D) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 in low-[Ca 2+ ] buffer in rat cerebrocortical NPMV fractions (n = 3; P = 0.49). Low-[Ca 2+ ] buffer contained 145 mM NaCl, 5 mM KCl, 20 mM MgCl 2 , 10 mM glucose and 20 mM <t>HEPES</t> (pH 7.5).
    Hepes Buffered Acsf, supplied by Tocris, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Tocris bath contained hepes buffered tyrode s solution
    Glu-induced GABA release is not mediated by vesicular release. (A) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in the presence of glutamate receptor antagonists (n = 7; P = 0.014; 128.5±13.4% of pre-stimulus control). (B) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 and Glu receptor antagonists in rat cerebrocortical <t>NPMV</t> fractions (n = 4; P = 0.82) (C) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in low-[Ca 2+ ]/high-[Mg 2+ ] buffer (n = 5; P = 0.91; 148.3±15.5% of pre-stimulus control). (D) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 in low-[Ca 2+ ] buffer in rat cerebrocortical NPMV fractions (n = 3; P = 0.49). Low-[Ca 2+ ] buffer contained 145 mM NaCl, 5 mM KCl, 20 mM MgCl 2 , 10 mM glucose and 20 mM <t>HEPES</t> (pH 7.5).
    Bath Contained Hepes Buffered Tyrode S Solution, supplied by Tocris, 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/bath contained hepes buffered tyrode s solution/product/Tocris
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    95
    Tocris hepes buffered saline
    Glu-induced GABA release is not mediated by vesicular release. (A) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in the presence of glutamate receptor antagonists (n = 7; P = 0.014; 128.5±13.4% of pre-stimulus control). (B) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 and Glu receptor antagonists in rat cerebrocortical <t>NPMV</t> fractions (n = 4; P = 0.82) (C) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in low-[Ca 2+ ]/high-[Mg 2+ ] buffer (n = 5; P = 0.91; 148.3±15.5% of pre-stimulus control). (D) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 in low-[Ca 2+ ] buffer in rat cerebrocortical NPMV fractions (n = 3; P = 0.49). Low-[Ca 2+ ] buffer contained 145 mM NaCl, 5 mM KCl, 20 mM MgCl 2 , 10 mM glucose and 20 mM <t>HEPES</t> (pH 7.5).
    Hepes Buffered Saline, supplied by Tocris, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    hepes buffered saline - by Bioz Stars, 2020-04
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    Glu-induced GABA release is not mediated by vesicular release. (A) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in the presence of glutamate receptor antagonists (n = 7; P = 0.014; 128.5±13.4% of pre-stimulus control). (B) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 and Glu receptor antagonists in rat cerebrocortical NPMV fractions (n = 4; P = 0.82) (C) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in low-[Ca 2+ ]/high-[Mg 2+ ] buffer (n = 5; P = 0.91; 148.3±15.5% of pre-stimulus control). (D) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 in low-[Ca 2+ ] buffer in rat cerebrocortical NPMV fractions (n = 3; P = 0.49). Low-[Ca 2+ ] buffer contained 145 mM NaCl, 5 mM KCl, 20 mM MgCl 2 , 10 mM glucose and 20 mM HEPES (pH 7.5).

    Journal: PLoS ONE

    Article Title: Glutamate Uptake Triggers Transporter-Mediated GABA Release from Astrocytes

    doi: 10.1371/journal.pone.0007153

    Figure Lengend Snippet: Glu-induced GABA release is not mediated by vesicular release. (A) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in the presence of glutamate receptor antagonists (n = 7; P = 0.014; 128.5±13.4% of pre-stimulus control). (B) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 and Glu receptor antagonists in rat cerebrocortical NPMV fractions (n = 4; P = 0.82) (C) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in low-[Ca 2+ ]/high-[Mg 2+ ] buffer (n = 5; P = 0.91; 148.3±15.5% of pre-stimulus control). (D) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 in low-[Ca 2+ ] buffer in rat cerebrocortical NPMV fractions (n = 3; P = 0.49). Low-[Ca 2+ ] buffer contained 145 mM NaCl, 5 mM KCl, 20 mM MgCl 2 , 10 mM glucose and 20 mM HEPES (pH 7.5).

    Article Snippet: Uptake studies were performed as follows: aliquots of NPMV fractions in HEPES buffer were preincubated with test compounds in the presence of NNC-711 (100 µM, Tocris) at 30°C for 10 min.

    Techniques: