Structured Review

Dojindo Labs hepes
Activity of β-GlcATs in microsomes from radish primary roots. a The microsomes (protein, 50 μg) were incubated under standard assay conditions with β-(1 → 3)-galactan as exogenous acceptor ( closed circles ) and with endogenous acceptors ( open circles ) for varying times up to 240 min. After 240 min, the amount of [ 14 C]GlcA transferred to β-(1 → 3)-galactan was about 0.6 % of initial UDP-[ 14 C]GlcA. The data are the mean + SE of quadruple experiments. b Relationship between enzyme activity and protein concentration. Various amounts of microsomal protein were incubated under standard conditions for 60 min. c Effects of pH on enzyme activity. Activity–pH curves result from experiments with buffers (80 mM) having different pH values under standard conditions for 120 min. Closed circles <t>Mes–KOH,</t> open circles <t>Hepes–KOH,</t> closed triangles acetate. d Activity–temperature curves result from incubations at different temperatures under standard assay conditions for 120 min. Note that the activity plotted for β-(1 → 3)-galactan includes that for endogenous acceptors in a and b
Hepes, supplied by Dojindo Labs, used in various techniques. Bioz Stars score: 95/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "Biosynthesis of the carbohydrate moieties of arabinogalactan proteins by membrane-bound ?-glucuronosyltransferases from radish primary roots"

Article Title: Biosynthesis of the carbohydrate moieties of arabinogalactan proteins by membrane-bound ?-glucuronosyltransferases from radish primary roots

Journal: Planta

doi: 10.1007/s00425-013-1959-0

Activity of β-GlcATs in microsomes from radish primary roots. a The microsomes (protein, 50 μg) were incubated under standard assay conditions with β-(1 → 3)-galactan as exogenous acceptor ( closed circles ) and with endogenous acceptors ( open circles ) for varying times up to 240 min. After 240 min, the amount of [ 14 C]GlcA transferred to β-(1 → 3)-galactan was about 0.6 % of initial UDP-[ 14 C]GlcA. The data are the mean + SE of quadruple experiments. b Relationship between enzyme activity and protein concentration. Various amounts of microsomal protein were incubated under standard conditions for 60 min. c Effects of pH on enzyme activity. Activity–pH curves result from experiments with buffers (80 mM) having different pH values under standard conditions for 120 min. Closed circles Mes–KOH, open circles Hepes–KOH, closed triangles acetate. d Activity–temperature curves result from incubations at different temperatures under standard assay conditions for 120 min. Note that the activity plotted for β-(1 → 3)-galactan includes that for endogenous acceptors in a and b
Figure Legend Snippet: Activity of β-GlcATs in microsomes from radish primary roots. a The microsomes (protein, 50 μg) were incubated under standard assay conditions with β-(1 → 3)-galactan as exogenous acceptor ( closed circles ) and with endogenous acceptors ( open circles ) for varying times up to 240 min. After 240 min, the amount of [ 14 C]GlcA transferred to β-(1 → 3)-galactan was about 0.6 % of initial UDP-[ 14 C]GlcA. The data are the mean + SE of quadruple experiments. b Relationship between enzyme activity and protein concentration. Various amounts of microsomal protein were incubated under standard conditions for 60 min. c Effects of pH on enzyme activity. Activity–pH curves result from experiments with buffers (80 mM) having different pH values under standard conditions for 120 min. Closed circles Mes–KOH, open circles Hepes–KOH, closed triangles acetate. d Activity–temperature curves result from incubations at different temperatures under standard assay conditions for 120 min. Note that the activity plotted for β-(1 → 3)-galactan includes that for endogenous acceptors in a and b

Techniques Used: Activity Assay, Incubation, Protein Concentration

2) Product Images from "Cross talk between polysulfide and nitric oxide in rat peritoneal mast cells"

Article Title: Cross talk between polysulfide and nitric oxide in rat peritoneal mast cells

Journal: American Journal of Physiology - Cell Physiology

doi: 10.1152/ajpcell.00028.2016

Effect of compound 48/80, Na 2 S 4 , GYY4137, and DEA NONOate on mast cell degranulation. A : translucent ( a–d ) and sulforhodamine-B (SFRM-B)-loaded fluorescent images of mast cells before ( e–h ) and after ( i–l ) stimulation with compound 48/80 for 10 min (1 μg/ml), Na 2 S 4 , GYY4137, and DEA NONOate for 20 min (50 μM each) in standard HEPES-buffered solution. Appearance of red fluorescence represents occurrence of exocytosis. Scale bar = 10 μm B : An example of time course changes of SFRM-B fluorescence intensity in mast cells stimulated with compound 48/80 (1 μg/ml) for 10 min in standard HEPES-buffered solution.
Figure Legend Snippet: Effect of compound 48/80, Na 2 S 4 , GYY4137, and DEA NONOate on mast cell degranulation. A : translucent ( a–d ) and sulforhodamine-B (SFRM-B)-loaded fluorescent images of mast cells before ( e–h ) and after ( i–l ) stimulation with compound 48/80 for 10 min (1 μg/ml), Na 2 S 4 , GYY4137, and DEA NONOate for 20 min (50 μM each) in standard HEPES-buffered solution. Appearance of red fluorescence represents occurrence of exocytosis. Scale bar = 10 μm B : An example of time course changes of SFRM-B fluorescence intensity in mast cells stimulated with compound 48/80 (1 μg/ml) for 10 min in standard HEPES-buffered solution.

Techniques Used: Fluorescence

Effect of Na 2 S 4 on intracellular Ca 2+ concentration ([Ca 2+ ] i ) in mast cells. A : a representative tracing showing temporal changes in fluo-4 fluorescence intensity (FI) induced by 50 μM Na 2 S 4 in the presence (solid line) or absence (dashed line) of extracellular Ca 2+ or in the absence of extracellular Ca 2+ but presence of ruthenium red (RR; 30 μM, dotted line). Changes in FI are shown by the percentage against the prestimulation intensity. In the presence of extracellular Ca 2+ , the cells were perfused with standard HEPES-buffered solution containing 1.3 mM CaCl 2 throughout the experiments, and Na 2 S 4 was applied after 2 min. In the absence of extracellular Ca 2+ , CaCl 2 was omitted. Instead, 1 mM EGTA was added, and the cells were perfused with the nominally Ca 2+ -free buffer for 3 min before starting image acquisition and throughout the experiments. B : dose-response relationship of Na 2 S 4 -induced changes in mean (±SE) summed area of fluorescence changes (SFC) in the presence (black bars; n = 12–62) or absence (hatched bars; n = 34–117) of extracellular Ca 2+ . P
Figure Legend Snippet: Effect of Na 2 S 4 on intracellular Ca 2+ concentration ([Ca 2+ ] i ) in mast cells. A : a representative tracing showing temporal changes in fluo-4 fluorescence intensity (FI) induced by 50 μM Na 2 S 4 in the presence (solid line) or absence (dashed line) of extracellular Ca 2+ or in the absence of extracellular Ca 2+ but presence of ruthenium red (RR; 30 μM, dotted line). Changes in FI are shown by the percentage against the prestimulation intensity. In the presence of extracellular Ca 2+ , the cells were perfused with standard HEPES-buffered solution containing 1.3 mM CaCl 2 throughout the experiments, and Na 2 S 4 was applied after 2 min. In the absence of extracellular Ca 2+ , CaCl 2 was omitted. Instead, 1 mM EGTA was added, and the cells were perfused with the nominally Ca 2+ -free buffer for 3 min before starting image acquisition and throughout the experiments. B : dose-response relationship of Na 2 S 4 -induced changes in mean (±SE) summed area of fluorescence changes (SFC) in the presence (black bars; n = 12–62) or absence (hatched bars; n = 34–117) of extracellular Ca 2+ . P

Techniques Used: Concentration Assay, Fluorescence

Effect of GYY4137 and diethylamine (DEA) NONOate on polysulfide production. A and C : typical examples of temporal changes in SSP4 FI induced by 50 μM of GYY4137 or DEA NONOate. Cells were perfused with standard HEPES-buffered solution throughout the experiments. B and D : dose-response relationships of GYY4137- and DEA NONOate-induced changes in SSP4 FI ( n = 9–25 and 19–38, respectively). Each column represents mean of summed area of fluorescence changes (SFC) ± SE. P
Figure Legend Snippet: Effect of GYY4137 and diethylamine (DEA) NONOate on polysulfide production. A and C : typical examples of temporal changes in SSP4 FI induced by 50 μM of GYY4137 or DEA NONOate. Cells were perfused with standard HEPES-buffered solution throughout the experiments. B and D : dose-response relationships of GYY4137- and DEA NONOate-induced changes in SSP4 FI ( n = 9–25 and 19–38, respectively). Each column represents mean of summed area of fluorescence changes (SFC) ± SE. P

Techniques Used: Fluorescence

Related Articles

Acetylene Reduction Assay:

Article Title: Biosynthesis of the carbohydrate moieties of arabinogalactan proteins by membrane-bound ?-glucuronosyltransferases from radish primary roots
Article Snippet: EGTA, Hepes, and Mes were from Dojindo Laboratories (Kumamoto, Japan). β-(1 → 3)- and β-(1 → 6)-Galactobioses, and -trioses were prepared from larch wood AG by partial acid hydrolysis (Aspinall et al. ). .. The α-l -Araf -(1 → 3)-β-Gal-(1 → 6)-Gal (l -Ara·Gal·Gal) was prepared by enzymatic hydrolysis of the Smith degradation product of acacia gum with exo-β-(1 → 3)-galactanase (Tsumuraya et al. ).

In Vitro:

Article Title: Production of Middle White Piglets after Transfer of Embryos Produced In Vitro
Article Snippet: Oocyte collection, in vitro maturation and fertilization were carried out as described previously [ , ]. .. All follicles were opened with a surgical blade in Medium 199 (with Hanks’ salts; Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% (v/v) fetal bovine serum (Gibco, Life Technologies, Carlsbad, CA, USA), 20 mM HEPES (Dojindo Laboratories, Kumamoto, Japan) and antibiotics (100 IU/ml penicillin G potassium (Sigma-Aldrich) and 0.1 mg/ml streptomycin sulfate (Sigma-Aldrich)).

MTT Assay:

Article Title: Yokukansan, a Traditional Japanese Medicine, Enhances the Glutamate Transporter GLT-1 Function in Cultured Rat Cortical Astrocytes
Article Snippet: .. EDTA, HEPES, and MTT were purchased from Dojindo (Kumamoto, Japan). .. Purified rabbit polyclonal antiserum to glial fibrillary acidic protein (GFAP) was purchased from Chemicon (Temecula, CA, USA).

Synthesized:

Article Title: Biosynthesis of the carbohydrate moieties of arabinogalactan proteins by membrane-bound ?-glucuronosyltransferases from radish primary roots
Article Snippet: EGTA, Hepes, and Mes were from Dojindo Laboratories (Kumamoto, Japan). β-(1 → 3)- and β-(1 → 6)-Galactobioses, and -trioses were prepared from larch wood AG by partial acid hydrolysis (Aspinall et al. ). .. Chemically synthesized β-(1 → 6)-galactopentaose was donated by Dr. Miura, Gifu Pharmaceutical University, Japan, and Professor Inazu, Tokai University, Japan (Miura et al. ). β-GlcA-(1 → 6)-Gal and β-GlcA-(1 → 6)-β-Gal-(1 → 6)-Gal were prepared from acacia gum (Kuroyama et al. ).

Protease Inhibitor:

Article Title: Yokukansan, a Traditional Japanese Medicine, Enhances the Glutamate Transporter GLT-1 Function in Cultured Rat Cortical Astrocytes
Article Snippet: EDTA, HEPES, and MTT were purchased from Dojindo (Kumamoto, Japan). .. For Western blotting, a protease inhibitor cocktail, blocking buffer (SuperBlock), and Can Get Signal Solutions were purchased from Sigma-Aldrich (St. Louis, MO, USA), Thermo Fisher Scientific (Rockford, IL, USA), and Toyobo (Osaka, Japan), respectively.

Cell Culture:

Article Title: Yokukansan, a Traditional Japanese Medicine, Enhances the Glutamate Transporter GLT-1 Function in Cultured Rat Cortical Astrocytes
Article Snippet: Reagents used in the cell culture experiments, dehydrokainate (DHK), pyrithiamine hydrobromide, Dulbecco's modified Eagle's medium (DMEM), DNase, glutamate dehydrogenase, β -nicotinamide adenine dinucleotide, 1-methoxyphenazine methosulphate, Triton X-100, TNF- α , and dBcAMP were purchased from Sigma-Aldrich (St. Louis, MO, USA). .. EDTA, HEPES, and MTT were purchased from Dojindo (Kumamoto, Japan).

Article Title: Production of Middle White Piglets after Transfer of Embryos Produced In Vitro
Article Snippet: All follicles were opened with a surgical blade in Medium 199 (with Hanks’ salts; Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% (v/v) fetal bovine serum (Gibco, Life Technologies, Carlsbad, CA, USA), 20 mM HEPES (Dojindo Laboratories, Kumamoto, Japan) and antibiotics (100 IU/ml penicillin G potassium (Sigma-Aldrich) and 0.1 mg/ml streptomycin sulfate (Sigma-Aldrich)). .. Approximately 20 COCs were cultured in 100 µl of maturation medium in 35-mm plastic dishes (TC dish 35 × 10, Nalge Nunc International, Roskilde, Denmark) for 20−22 h. They were then cultured in each medium without dbcAMP and hormones for 24 h. Maturation culture was carried out at 39 C under CO2 and N2 adjusted to 5% and 90%, respectively (5% O2 ).

Purification:

Article Title: Escherichia coli Cytosolic Glycerophosphodiester Phosphodiesterase (UgpQ) Requires Mg2+, Co2+, or Mn2+ for Its Enzyme Activity ▿
Article Snippet: Anti-UgpQ rabbit polyclonal antibody was made by Hokudo Co., Ltd. (Hokkaido, Japan), using purified UgpQ as an antigen. .. MES (morpholineethanesulfonic acid), PIPES [piperazine- N , N ′-bis(2-ethanesulfonic acid)], MOPS (morpholinepropanesulfonic acid), HEPES, TAPS [ N -tris(hydroxymethyl)methyl-3-amino-propanesulfonic acid], and CHES [2-(cyclohexylamino)ethanesulfonic acid] were purchased from Dojindo (Kumamoto, Japan).

Article Title: Yokukansan, a Traditional Japanese Medicine, Enhances the Glutamate Transporter GLT-1 Function in Cultured Rat Cortical Astrocytes
Article Snippet: EDTA, HEPES, and MTT were purchased from Dojindo (Kumamoto, Japan). .. Purified rabbit polyclonal antiserum to glial fibrillary acidic protein (GFAP) was purchased from Chemicon (Temecula, CA, USA).

Polymerase Chain Reaction:

Article Title: Escherichia coli Cytosolic Glycerophosphodiester Phosphodiesterase (UgpQ) Requires Mg2+, Co2+, or Mn2+ for Its Enzyme Activity ▿
Article Snippet: PCR primers were purchased from Operon Biotechnologies (Tokyo, Japan). .. MES (morpholineethanesulfonic acid), PIPES [piperazine- N , N ′-bis(2-ethanesulfonic acid)], MOPS (morpholinepropanesulfonic acid), HEPES, TAPS [ N -tris(hydroxymethyl)methyl-3-amino-propanesulfonic acid], and CHES [2-(cyclohexylamino)ethanesulfonic acid] were purchased from Dojindo (Kumamoto, Japan).

other:

Article Title: Electrophysiological measurement of ion channels on plasma/organelle membranes using an on-chip lipid bilayer system
Article Snippet: HEPES was purchased from Dojindo Molecular Technologies (Kumamoto, Japan), Verapamil was purchased from Sigma-Aldrich (St. Louis USA), and astemizole was purchased from Toronto Research Chemicals (North York, Canada).

Article Title: Sucrose assists selection of high‐quality oocytes in pigs, et al. Sucrose assists selection of high‐quality oocytes in pigs
Article Snippet: 2.2 Sucrose treatment After IVM, cumulus cells were removed from the COCs by gentle pipetting after treatment with 0.1% (w/v) hyaluronidase (Sigma) for 2–5 min, and the denuded oocytes were then washed three times in Medium 199 (with Hanks’ balanced salts; Sigma) supplemented with 10% (v/v) fetal calf serum (FCS) (Gibco, Life Technologies Inc., Grand Island, NE, USA), 20 mmol/L HEPES (Dojindo Laboratories, Kumamoto, Japan), antibiotics (100 units/mL penicillin G potassium [Sigma] and 0.1 mg/mL streptomycin sulfate [Sigma]), pH adjusted to 7.4.

Article Title: ATP7B expression confers multidrug resistance through drug sequestration
Article Snippet: Chemicals We purchased G-418, cisplatin, doxorubicin etoposide, paclitaxel and tamoxifen form Sigma Chemical Co., (St, Louis, MO), Glutamine, NH4 Cl and CuCl2 from Wako (Osaka, Japan), and HEPES and BCS from Dojindo (Kumamoto, Japan).

Article Title: Cross talk between polysulfide and nitric oxide in rat peritoneal mast cells
Article Snippet: GYY4137 [morpholin-4-ium 4 methoxphenyl (morpholino) phosphinodithionate], fluo-4/AM, HEPES, N G -monomethyl- l -arginine ( l -NMMA), carboxy-PTIO (cPTIO), and SSP4 [sulfane sulfur probe 4, 3′,6′-di( O -thiosalicyl)fluorescein] were obtained from Dojindo (Kumamoto, Japan).

Blocking Assay:

Article Title: Yokukansan, a Traditional Japanese Medicine, Enhances the Glutamate Transporter GLT-1 Function in Cultured Rat Cortical Astrocytes
Article Snippet: EDTA, HEPES, and MTT were purchased from Dojindo (Kumamoto, Japan). .. For Western blotting, a protease inhibitor cocktail, blocking buffer (SuperBlock), and Can Get Signal Solutions were purchased from Sigma-Aldrich (St. Louis, MO, USA), Thermo Fisher Scientific (Rockford, IL, USA), and Toyobo (Osaka, Japan), respectively.

Modification:

Article Title: Yokukansan, a Traditional Japanese Medicine, Enhances the Glutamate Transporter GLT-1 Function in Cultured Rat Cortical Astrocytes
Article Snippet: Reagents used in the cell culture experiments, dehydrokainate (DHK), pyrithiamine hydrobromide, Dulbecco's modified Eagle's medium (DMEM), DNase, glutamate dehydrogenase, β -nicotinamide adenine dinucleotide, 1-methoxyphenazine methosulphate, Triton X-100, TNF- α , and dBcAMP were purchased from Sigma-Aldrich (St. Louis, MO, USA). .. EDTA, HEPES, and MTT were purchased from Dojindo (Kumamoto, Japan).

Article Title: Effects of pantothenic acid supplement on secretion of steroids by the adrenal cortex in female rats
Article Snippet: .. Dulbecco's modified Eagle's medium (DMEM), MEM non‐essential amino acids solution, penicillin and streptomycin (Invitrogen, Burlington, ON, Canada), HEPES (Dojindo, Gaitherburg, MD, USA), collagenase type V, deoxyribonuclease, fetal bovine serum (Sigma–Aldrich, St Louis, MO, USA), rat adrenocorticotropic hormone (ACTH) (AFPRFR7890, NIDDK, Torrance, CA, USA), PaA (calcium pantothenate) (Daiichi Fine Chemical, Tokyo, Japan), and aprotinin (Sigma–Aldrich) were purchased. .. Wistar Imamichi rats from the Imamichi Institute for Animal Reproduction (Kasumigaura, Ibaraki, Japan) were used in this study.

Article Title: Generation of Live Piglets from Cryopreserved Oocytes for the First Time Using a Defined System for In Vitro Embryo Production
Article Snippet: Ovaries from prepubertal cross-bred gilts (Landrace, Large White, and Duroc breeds) were collected at a local slaughterhouse and transported to the laboratory in Dulbecco’s phosphate-buffered saline (PBS) (Nissui Pharmaceutical Co. Ltd, Tokyo, Japan) at 35 to 37°C within 1 h. Cumulus–oocyte complexes (COCs) were collected by scraping of 3- to 6-mm follicles into a collection medium consisting of Medium 199 (with Hanks’ salts) supplemented with 5% (v/v) fetal bovine serum (Gibco; Invitrogen Corp., Carlsbad, CA, USA), 20 mM HEPES (Dojindo Laboratories, Kumamoto, Japan), and antibiotics [100 units/ml penicillin G potassium and 0.1 mg/ml streptomycin sulfate]. .. In brief, COCs were treated for 30 min in a basic medium (BM) consisting of modified NCSU-37 without glucose, but supplemented with 20 mM HEPES, 50 µM β-mercaptoethanol, 0.17 mM sodium pyruvate, 2.73 mM sodium lactate.

Article Title: Lysine suppresses myofibrillar protein degradation by regulating the autophagic-lysosomal system through phosphorylation of Akt in C2C12 cells
Article Snippet: Dulbecco’s modified Eagle’s medium (DMEM, low glucose), minimum essential medium (MEM) Vitamin mix, Akti and dimethyl sulfoxide (DMSO) were obtained from Sigma (St. Louis, MO, USA). .. HEPES was obtained from Dojindo Molecular Technologies, Inc. (Kumamoto, Japan).

Western Blot:

Article Title: Yokukansan, a Traditional Japanese Medicine, Enhances the Glutamate Transporter GLT-1 Function in Cultured Rat Cortical Astrocytes
Article Snippet: EDTA, HEPES, and MTT were purchased from Dojindo (Kumamoto, Japan). .. For Western blotting, a protease inhibitor cocktail, blocking buffer (SuperBlock), and Can Get Signal Solutions were purchased from Sigma-Aldrich (St. Louis, MO, USA), Thermo Fisher Scientific (Rockford, IL, USA), and Toyobo (Osaka, Japan), respectively.

Plasmid Preparation:

Article Title: Yokukansan, a Traditional Japanese Medicine, Enhances the Glutamate Transporter GLT-1 Function in Cultured Rat Cortical Astrocytes
Article Snippet: EDTA, HEPES, and MTT were purchased from Dojindo (Kumamoto, Japan). .. EDTA, HEPES, and MTT were purchased from Dojindo (Kumamoto, Japan).

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    Dojindo Labs ca2 hepes tyrode s solution
    Time course of FRET signals of various α-actinin–YC-Nanos. (A) Time course of changes in F yellow / F cyan during spontaneous beating in myocytes expressing α-actinin–YC-Nano15, α-actinin–YC-Nano50, α-actinin–YC-Nano65, or α-actinin–YC-Nano140 in the Z disks. n = 4–12 cells. For strong possible fluorescence signal recordings, measurements were performed in 2.0 mM Ca 2+ <t>-HEPES–Tyrode’s</t> solution at 37°C. See Video 2. (B) Maximal ( R max ) and minimal ( R 0 ) FRET signals for α-actinin–YC-Nano15, α-actinin–YC-Nano50, α-actinin–YC-Nano65, and α-actinin–YC-Nano140 expressed in the Z disks of myocytes during spontaneous beating (data obtained for 5 s as in A). Closed symbols, R 0 ; open symbols, R max . (C) Values of Δ R (i.e., R max − R 0 ) for α-actinin–YC-Nanos. Data obtained from A and B. No significant differences were observed between groups (Tukey-Kramer test). (D) Values of the ratio of Δ R to R 0 (i.e., Δ R / R 0 ) for α-actinin–YC-Nanos. **, P
    Ca2 Hepes Tyrode S Solution, supplied by Dojindo Labs, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ca2 hepes tyrode s solution/product/Dojindo Labs
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    Dojindo Labs hepes
    Activity of β-GlcATs in microsomes from radish primary roots. a The microsomes (protein, 50 μg) were incubated under standard assay conditions with β-(1 → 3)-galactan as exogenous acceptor ( closed circles ) and with endogenous acceptors ( open circles ) for varying times up to 240 min. After 240 min, the amount of [ 14 C]GlcA transferred to β-(1 → 3)-galactan was about 0.6 % of initial UDP-[ 14 C]GlcA. The data are the mean + SE of quadruple experiments. b Relationship between enzyme activity and protein concentration. Various amounts of microsomal protein were incubated under standard conditions for 60 min. c Effects of pH on enzyme activity. Activity–pH curves result from experiments with buffers (80 mM) having different pH values under standard conditions for 120 min. Closed circles <t>Mes–KOH,</t> open circles <t>Hepes–KOH,</t> closed triangles acetate. d Activity–temperature curves result from incubations at different temperatures under standard assay conditions for 120 min. Note that the activity plotted for β-(1 → 3)-galactan includes that for endogenous acceptors in a and b
    Hepes, supplied by Dojindo Labs, used in various techniques. Bioz Stars score: 95/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes/product/Dojindo Labs
    Average 95 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    hepes - by Bioz Stars, 2020-04
    95/100 stars
      Buy from Supplier

    91
    Dojindo Labs buffers hepes
    Activity of β-GlcATs in microsomes from radish primary roots. a The microsomes (protein, 50 μg) were incubated under standard assay conditions with β-(1 → 3)-galactan as exogenous acceptor ( closed circles ) and with endogenous acceptors ( open circles ) for varying times up to 240 min. After 240 min, the amount of [ 14 C]GlcA transferred to β-(1 → 3)-galactan was about 0.6 % of initial UDP-[ 14 C]GlcA. The data are the mean + SE of quadruple experiments. b Relationship between enzyme activity and protein concentration. Various amounts of microsomal protein were incubated under standard conditions for 60 min. c Effects of pH on enzyme activity. Activity–pH curves result from experiments with buffers (80 mM) having different pH values under standard conditions for 120 min. Closed circles <t>Mes–KOH,</t> open circles <t>Hepes–KOH,</t> closed triangles acetate. d Activity–temperature curves result from incubations at different temperatures under standard assay conditions for 120 min. Note that the activity plotted for β-(1 → 3)-galactan includes that for endogenous acceptors in a and b
    Buffers Hepes, supplied by Dojindo Labs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/buffers hepes/product/Dojindo Labs
    Average 91 stars, based on 1 article reviews
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    Time course of FRET signals of various α-actinin–YC-Nanos. (A) Time course of changes in F yellow / F cyan during spontaneous beating in myocytes expressing α-actinin–YC-Nano15, α-actinin–YC-Nano50, α-actinin–YC-Nano65, or α-actinin–YC-Nano140 in the Z disks. n = 4–12 cells. For strong possible fluorescence signal recordings, measurements were performed in 2.0 mM Ca 2+ -HEPES–Tyrode’s solution at 37°C. See Video 2. (B) Maximal ( R max ) and minimal ( R 0 ) FRET signals for α-actinin–YC-Nano15, α-actinin–YC-Nano50, α-actinin–YC-Nano65, and α-actinin–YC-Nano140 expressed in the Z disks of myocytes during spontaneous beating (data obtained for 5 s as in A). Closed symbols, R 0 ; open symbols, R max . (C) Values of Δ R (i.e., R max − R 0 ) for α-actinin–YC-Nanos. Data obtained from A and B. No significant differences were observed between groups (Tukey-Kramer test). (D) Values of the ratio of Δ R to R 0 (i.e., Δ R / R 0 ) for α-actinin–YC-Nanos. **, P

    Journal: The Journal of General Physiology

    Article Title: Simultaneous imaging of local calcium and single sarcomere length in rat neonatal cardiomyocytes using yellow Cameleon-Nano140

    doi: 10.1085/jgp.201611604

    Figure Lengend Snippet: Time course of FRET signals of various α-actinin–YC-Nanos. (A) Time course of changes in F yellow / F cyan during spontaneous beating in myocytes expressing α-actinin–YC-Nano15, α-actinin–YC-Nano50, α-actinin–YC-Nano65, or α-actinin–YC-Nano140 in the Z disks. n = 4–12 cells. For strong possible fluorescence signal recordings, measurements were performed in 2.0 mM Ca 2+ -HEPES–Tyrode’s solution at 37°C. See Video 2. (B) Maximal ( R max ) and minimal ( R 0 ) FRET signals for α-actinin–YC-Nano15, α-actinin–YC-Nano50, α-actinin–YC-Nano65, and α-actinin–YC-Nano140 expressed in the Z disks of myocytes during spontaneous beating (data obtained for 5 s as in A). Closed symbols, R 0 ; open symbols, R max . (C) Values of Δ R (i.e., R max − R 0 ) for α-actinin–YC-Nanos. Data obtained from A and B. No significant differences were observed between groups (Tukey-Kramer test). (D) Values of the ratio of Δ R to R 0 (i.e., Δ R / R 0 ) for α-actinin–YC-Nanos. **, P

    Article Snippet: Experimental procedures For Ca2+ imaging with Fluo-4, α-actinin–YC-Nano140–expressing cardiomyocytes were preincubated in 1.5 mM Ca2+ -HEPES–Tyrode’s solution containing 2 µM Fluo-4-AM (Dojindo) for 20 min at 25°C.

    Techniques: Expressing, Fluorescence

    Activity of β-GlcATs in microsomes from radish primary roots. a The microsomes (protein, 50 μg) were incubated under standard assay conditions with β-(1 → 3)-galactan as exogenous acceptor ( closed circles ) and with endogenous acceptors ( open circles ) for varying times up to 240 min. After 240 min, the amount of [ 14 C]GlcA transferred to β-(1 → 3)-galactan was about 0.6 % of initial UDP-[ 14 C]GlcA. The data are the mean + SE of quadruple experiments. b Relationship between enzyme activity and protein concentration. Various amounts of microsomal protein were incubated under standard conditions for 60 min. c Effects of pH on enzyme activity. Activity–pH curves result from experiments with buffers (80 mM) having different pH values under standard conditions for 120 min. Closed circles Mes–KOH, open circles Hepes–KOH, closed triangles acetate. d Activity–temperature curves result from incubations at different temperatures under standard assay conditions for 120 min. Note that the activity plotted for β-(1 → 3)-galactan includes that for endogenous acceptors in a and b

    Journal: Planta

    Article Title: Biosynthesis of the carbohydrate moieties of arabinogalactan proteins by membrane-bound ?-glucuronosyltransferases from radish primary roots

    doi: 10.1007/s00425-013-1959-0

    Figure Lengend Snippet: Activity of β-GlcATs in microsomes from radish primary roots. a The microsomes (protein, 50 μg) were incubated under standard assay conditions with β-(1 → 3)-galactan as exogenous acceptor ( closed circles ) and with endogenous acceptors ( open circles ) for varying times up to 240 min. After 240 min, the amount of [ 14 C]GlcA transferred to β-(1 → 3)-galactan was about 0.6 % of initial UDP-[ 14 C]GlcA. The data are the mean + SE of quadruple experiments. b Relationship between enzyme activity and protein concentration. Various amounts of microsomal protein were incubated under standard conditions for 60 min. c Effects of pH on enzyme activity. Activity–pH curves result from experiments with buffers (80 mM) having different pH values under standard conditions for 120 min. Closed circles Mes–KOH, open circles Hepes–KOH, closed triangles acetate. d Activity–temperature curves result from incubations at different temperatures under standard assay conditions for 120 min. Note that the activity plotted for β-(1 → 3)-galactan includes that for endogenous acceptors in a and b

    Article Snippet: EGTA, Hepes, and Mes were from Dojindo Laboratories (Kumamoto, Japan). β-(1 → 3)- and β-(1 → 6)-Galactobioses, and -trioses were prepared from larch wood AG by partial acid hydrolysis (Aspinall et al. ).

    Techniques: Activity Assay, Incubation, Protein Concentration

    Effect of compound 48/80, Na 2 S 4 , GYY4137, and DEA NONOate on mast cell degranulation. A : translucent ( a–d ) and sulforhodamine-B (SFRM-B)-loaded fluorescent images of mast cells before ( e–h ) and after ( i–l ) stimulation with compound 48/80 for 10 min (1 μg/ml), Na 2 S 4 , GYY4137, and DEA NONOate for 20 min (50 μM each) in standard HEPES-buffered solution. Appearance of red fluorescence represents occurrence of exocytosis. Scale bar = 10 μm B : An example of time course changes of SFRM-B fluorescence intensity in mast cells stimulated with compound 48/80 (1 μg/ml) for 10 min in standard HEPES-buffered solution.

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Cross talk between polysulfide and nitric oxide in rat peritoneal mast cells

    doi: 10.1152/ajpcell.00028.2016

    Figure Lengend Snippet: Effect of compound 48/80, Na 2 S 4 , GYY4137, and DEA NONOate on mast cell degranulation. A : translucent ( a–d ) and sulforhodamine-B (SFRM-B)-loaded fluorescent images of mast cells before ( e–h ) and after ( i–l ) stimulation with compound 48/80 for 10 min (1 μg/ml), Na 2 S 4 , GYY4137, and DEA NONOate for 20 min (50 μM each) in standard HEPES-buffered solution. Appearance of red fluorescence represents occurrence of exocytosis. Scale bar = 10 μm B : An example of time course changes of SFRM-B fluorescence intensity in mast cells stimulated with compound 48/80 (1 μg/ml) for 10 min in standard HEPES-buffered solution.

    Article Snippet: GYY4137 [morpholin-4-ium 4 methoxphenyl (morpholino) phosphinodithionate], fluo-4/AM, HEPES, N G -monomethyl- l -arginine ( l -NMMA), carboxy-PTIO (cPTIO), and SSP4 [sulfane sulfur probe 4, 3′,6′-di( O -thiosalicyl)fluorescein] were obtained from Dojindo (Kumamoto, Japan).

    Techniques: Fluorescence

    Effect of Na 2 S 4 on intracellular Ca 2+ concentration ([Ca 2+ ] i ) in mast cells. A : a representative tracing showing temporal changes in fluo-4 fluorescence intensity (FI) induced by 50 μM Na 2 S 4 in the presence (solid line) or absence (dashed line) of extracellular Ca 2+ or in the absence of extracellular Ca 2+ but presence of ruthenium red (RR; 30 μM, dotted line). Changes in FI are shown by the percentage against the prestimulation intensity. In the presence of extracellular Ca 2+ , the cells were perfused with standard HEPES-buffered solution containing 1.3 mM CaCl 2 throughout the experiments, and Na 2 S 4 was applied after 2 min. In the absence of extracellular Ca 2+ , CaCl 2 was omitted. Instead, 1 mM EGTA was added, and the cells were perfused with the nominally Ca 2+ -free buffer for 3 min before starting image acquisition and throughout the experiments. B : dose-response relationship of Na 2 S 4 -induced changes in mean (±SE) summed area of fluorescence changes (SFC) in the presence (black bars; n = 12–62) or absence (hatched bars; n = 34–117) of extracellular Ca 2+ . P

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Cross talk between polysulfide and nitric oxide in rat peritoneal mast cells

    doi: 10.1152/ajpcell.00028.2016

    Figure Lengend Snippet: Effect of Na 2 S 4 on intracellular Ca 2+ concentration ([Ca 2+ ] i ) in mast cells. A : a representative tracing showing temporal changes in fluo-4 fluorescence intensity (FI) induced by 50 μM Na 2 S 4 in the presence (solid line) or absence (dashed line) of extracellular Ca 2+ or in the absence of extracellular Ca 2+ but presence of ruthenium red (RR; 30 μM, dotted line). Changes in FI are shown by the percentage against the prestimulation intensity. In the presence of extracellular Ca 2+ , the cells were perfused with standard HEPES-buffered solution containing 1.3 mM CaCl 2 throughout the experiments, and Na 2 S 4 was applied after 2 min. In the absence of extracellular Ca 2+ , CaCl 2 was omitted. Instead, 1 mM EGTA was added, and the cells were perfused with the nominally Ca 2+ -free buffer for 3 min before starting image acquisition and throughout the experiments. B : dose-response relationship of Na 2 S 4 -induced changes in mean (±SE) summed area of fluorescence changes (SFC) in the presence (black bars; n = 12–62) or absence (hatched bars; n = 34–117) of extracellular Ca 2+ . P

    Article Snippet: GYY4137 [morpholin-4-ium 4 methoxphenyl (morpholino) phosphinodithionate], fluo-4/AM, HEPES, N G -monomethyl- l -arginine ( l -NMMA), carboxy-PTIO (cPTIO), and SSP4 [sulfane sulfur probe 4, 3′,6′-di( O -thiosalicyl)fluorescein] were obtained from Dojindo (Kumamoto, Japan).

    Techniques: Concentration Assay, Fluorescence

    Effect of GYY4137 and diethylamine (DEA) NONOate on polysulfide production. A and C : typical examples of temporal changes in SSP4 FI induced by 50 μM of GYY4137 or DEA NONOate. Cells were perfused with standard HEPES-buffered solution throughout the experiments. B and D : dose-response relationships of GYY4137- and DEA NONOate-induced changes in SSP4 FI ( n = 9–25 and 19–38, respectively). Each column represents mean of summed area of fluorescence changes (SFC) ± SE. P

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Cross talk between polysulfide and nitric oxide in rat peritoneal mast cells

    doi: 10.1152/ajpcell.00028.2016

    Figure Lengend Snippet: Effect of GYY4137 and diethylamine (DEA) NONOate on polysulfide production. A and C : typical examples of temporal changes in SSP4 FI induced by 50 μM of GYY4137 or DEA NONOate. Cells were perfused with standard HEPES-buffered solution throughout the experiments. B and D : dose-response relationships of GYY4137- and DEA NONOate-induced changes in SSP4 FI ( n = 9–25 and 19–38, respectively). Each column represents mean of summed area of fluorescence changes (SFC) ± SE. P

    Article Snippet: GYY4137 [morpholin-4-ium 4 methoxphenyl (morpholino) phosphinodithionate], fluo-4/AM, HEPES, N G -monomethyl- l -arginine ( l -NMMA), carboxy-PTIO (cPTIO), and SSP4 [sulfane sulfur probe 4, 3′,6′-di( O -thiosalicyl)fluorescein] were obtained from Dojindo (Kumamoto, Japan).

    Techniques: Fluorescence