2dog atp energy clamp  (Worthington Biochemical)


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    Structured Review

    Worthington Biochemical 2dog atp energy clamp
    The 2-deoxyglucose <t>(2DOG)</t> energy clamp. (A) Graphic depiction. Saturating amounts of 2-deoxyglucose (2DOG) and hexokinase (HK) recycle <t>ATP</t> back to ADP by rapidly and irreversibly converting 2DOG into 2-deoxyglucose phosphate (2DOGP). The ADP concentration is clamped at levels determined by the amount of ADP added. IMM = inner mitochondrial membrane. (B and C) Oxygraph tracings of inner membrane potential (inversely related to electrode potential shown on y-axis after calculation using the Nernst equation) vs. time obtained by incubating mouse hindlimb muscle mitochondria (mito), 0.05 mg/ml, fueled by 5 mM succinate (panel B) or 5 mM succinate plus 5 μM rotenone (panel C). ADP was added in incremental amounts to generate the final recycling nucleotide phosphate concentrations shown. After each addition, plateau values were reached, consistent with recycling at a steady [ADP].
    2dog Atp Energy Clamp, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 90/100, based on 78 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/2dog atp energy clamp/product/Worthington Biochemical
    Average 90 stars, based on 78 article reviews
    Price from $9.99 to $1999.99
    2dog atp energy clamp - by Bioz Stars, 2020-10
    90/100 stars

    Images

    1) Product Images from "Voltage-Dependent Regulation of Complex II Energized Mitochondrial Oxygen Flux"

    Article Title: Voltage-Dependent Regulation of Complex II Energized Mitochondrial Oxygen Flux

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0154982

    The 2-deoxyglucose (2DOG) energy clamp. (A) Graphic depiction. Saturating amounts of 2-deoxyglucose (2DOG) and hexokinase (HK) recycle ATP back to ADP by rapidly and irreversibly converting 2DOG into 2-deoxyglucose phosphate (2DOGP). The ADP concentration is clamped at levels determined by the amount of ADP added. IMM = inner mitochondrial membrane. (B and C) Oxygraph tracings of inner membrane potential (inversely related to electrode potential shown on y-axis after calculation using the Nernst equation) vs. time obtained by incubating mouse hindlimb muscle mitochondria (mito), 0.05 mg/ml, fueled by 5 mM succinate (panel B) or 5 mM succinate plus 5 μM rotenone (panel C). ADP was added in incremental amounts to generate the final recycling nucleotide phosphate concentrations shown. After each addition, plateau values were reached, consistent with recycling at a steady [ADP].
    Figure Legend Snippet: The 2-deoxyglucose (2DOG) energy clamp. (A) Graphic depiction. Saturating amounts of 2-deoxyglucose (2DOG) and hexokinase (HK) recycle ATP back to ADP by rapidly and irreversibly converting 2DOG into 2-deoxyglucose phosphate (2DOGP). The ADP concentration is clamped at levels determined by the amount of ADP added. IMM = inner mitochondrial membrane. (B and C) Oxygraph tracings of inner membrane potential (inversely related to electrode potential shown on y-axis after calculation using the Nernst equation) vs. time obtained by incubating mouse hindlimb muscle mitochondria (mito), 0.05 mg/ml, fueled by 5 mM succinate (panel B) or 5 mM succinate plus 5 μM rotenone (panel C). ADP was added in incremental amounts to generate the final recycling nucleotide phosphate concentrations shown. After each addition, plateau values were reached, consistent with recycling at a steady [ADP].

    Techniques Used: Concentration Assay

    2) Product Images from "Voltage-Dependent Regulation of Complex II Energized Mitochondrial Oxygen Flux"

    Article Title: Voltage-Dependent Regulation of Complex II Energized Mitochondrial Oxygen Flux

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0154982

    The 2-deoxyglucose (2DOG) energy clamp. (A) Graphic depiction. Saturating amounts of 2-deoxyglucose (2DOG) and hexokinase (HK) recycle ATP back to ADP by rapidly and irreversibly converting 2DOG into 2-deoxyglucose phosphate (2DOGP). The ADP concentration is clamped at levels determined by the amount of ADP added. IMM = inner mitochondrial membrane. (B and C) Oxygraph tracings of inner membrane potential (inversely related to electrode potential shown on y-axis after calculation using the Nernst equation) vs. time obtained by incubating mouse hindlimb muscle mitochondria (mito), 0.05 mg/ml, fueled by 5 mM succinate (panel B) or 5 mM succinate plus 5 μM rotenone (panel C). ADP was added in incremental amounts to generate the final recycling nucleotide phosphate concentrations shown. After each addition, plateau values were reached, consistent with recycling at a steady [ADP].
    Figure Legend Snippet: The 2-deoxyglucose (2DOG) energy clamp. (A) Graphic depiction. Saturating amounts of 2-deoxyglucose (2DOG) and hexokinase (HK) recycle ATP back to ADP by rapidly and irreversibly converting 2DOG into 2-deoxyglucose phosphate (2DOGP). The ADP concentration is clamped at levels determined by the amount of ADP added. IMM = inner mitochondrial membrane. (B and C) Oxygraph tracings of inner membrane potential (inversely related to electrode potential shown on y-axis after calculation using the Nernst equation) vs. time obtained by incubating mouse hindlimb muscle mitochondria (mito), 0.05 mg/ml, fueled by 5 mM succinate (panel B) or 5 mM succinate plus 5 μM rotenone (panel C). ADP was added in incremental amounts to generate the final recycling nucleotide phosphate concentrations shown. After each addition, plateau values were reached, consistent with recycling at a steady [ADP].

    Techniques Used: Concentration Assay

    Related Articles

    Isolation:

    Article Title: Voltage-Dependent Regulation of Complex II Energized Mitochondrial Oxygen Flux
    Article Snippet: .. Use of the 2DOG ATP energy clamp to quantify ATP production in isolated mitochondria and simultaneous assessment of H2 O2 production Mitochondria (0.1 mg/ml) were added to individual wells of black polystyrene 96-well round bottom plates in a total volume of 60 μl and incubated at 37°C in respiratory buffer plus 5 units/ml HK (Worthington Biochemical) and 5 mM 2DOG in the presence of selected concentrations of ADP. .. After incubation for 20 min with gentle rocking, the contents of the microplate wells were removed to tubes on ice containing 1 μl of 120 μM oligomycin to inhibit ATP synthase.

    Incubation:

    Article Title: Voltage-Dependent Regulation of Complex II Energized Mitochondrial Oxygen Flux
    Article Snippet: .. Use of the 2DOG ATP energy clamp to quantify ATP production in isolated mitochondria and simultaneous assessment of H2 O2 production Mitochondria (0.1 mg/ml) were added to individual wells of black polystyrene 96-well round bottom plates in a total volume of 60 μl and incubated at 37°C in respiratory buffer plus 5 units/ml HK (Worthington Biochemical) and 5 mM 2DOG in the presence of selected concentrations of ADP. .. After incubation for 20 min with gentle rocking, the contents of the microplate wells were removed to tubes on ice containing 1 μl of 120 μM oligomycin to inhibit ATP synthase.

    Article Title: Dietary fat, fatty acid saturation and mitochondrial bioenergetics
    Article Snippet: .. Mitochondria (0.5 mg/ml for liver or 0.1 mg/ml for heart) were added to individual wells of 96-well plates in a total volume of 60 μl and incubated at 37°C in respiratory buffer plus 5 units/ml HK (Worthington Biochemical) and 5 mM 2DOG in the presence of selected concentrations of ADP and fueled by the combined substrates, 5 mM succinate + 5 mM glutamate + 1 mM malate. .. After incubation for 20 min, the contents of the microplate wells were removed to tubes on ice containing 1μl of 120 μM oligomycin to inhibit ATP synthase.

    Article Title: Voltage-Dependent Regulation of Complex II Energized Mitochondrial Oxygen Flux
    Article Snippet: .. Mitochondria (0.05 mg/ml) were incubated at 37°C in 2 ml of ionic respiratory buffer (105 mM KCl, 10 mM NaCl, 5 mM Na2 HPO4 , 2 mM MgCl2 , 10 mM HEPES pH 7.2, 1 mM EGTA, 0.2% defatted BSA) with 5 U/ml hexokinase (Worthington Biochemical), and 5 mM 2-deoxyglucose. ..

    other:

    Article Title: Pharmacological Inhibition of GPR4 remediates intestinal inflammation in a mouse colitis model
    Article Snippet: [ ] [ ] [ ] Okada Y, Wu D, Trynka G, Raj T, Terao C, Ikari K, Kochi Y, Ohmura K, Suzuki A, Yoshida S, Graham RR, Manoharan A, Ortmann W, Bhangale T, Denny JC, Carroll RJ, Eyler AE, Greenberg JD, Kremer JM, Pappas DA, Jiang L, Yin J, Ye L, Su DF, Yang J, Xie G, Keystone E, Westra HJ, Esko T, Metspalu A, Zhou X, Gupta N, Mirel D, Stahl EA, Diogo D, Cui J, Liao K, Guo MH, Myouzen K, Kawaguchi T, Coenen MJ, van Riel PL, van de Laar MA, Guchelaar HJ, Huizinga TW, Dieude P, Mariette X, Bridges SL Jr., Zhernakova A, Toes RE, Tak PP, Miceli-Richard C, Bang SY, Lee HS, Martin J, Gonzalez-Gay MA, Rodriguez-Rodriguez L, Rantapaa-Dahlqvist S, Arlestig L, Choi HK, Kamatani Y, Galan P, Lathrop M, Eyre S, Bowes J, Barton A, de Vries N, Moreland LW, Criswell LA, Karlson EW, Taniguchi A, Yamada R, Kubo M, Liu JS, Bae SC, Worthington J, Padyukov L, Klareskog L, Gregersen PK, Raychaudhuri S, Stranger BE, De Jager PL, Franke L, Visscher PM, Brown MA, Yamanaka H, Mimori T, Takahashi A, Xu H, Behrens TW, Siminovitch KA, Momohara S, Matsuda F, Yamamoto K, Plenge RM, 2014.

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