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Ab109903

Manufactured by Abcam
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Ab109903 is a primary antibody targeting Myc-tag. It is designed for use in various immunological techniques such as Western blotting, immunoprecipitation, and immunocytochemistry.

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Lab products found in correlation

Ab109906, by Abcam (3 mentions) Ab109907, by Abcam (3 mentions) Ab109905, by Abcam (2 mentions) Xf96 extracellular flux analyzer, by Agilent Technologies (2 mentions) Citrate synthase activity assay kit, by Abcam (1 mentions)

5 protocols using ab109903

1

Metformin Effects on Hepatocyte Mitochondria

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Primary hepatocytes were seeded in an XF 96 well plate coated with 0.01% collagen type 1.36 h after seeding, cells were washed with PBS twice, and glucose production medium was added with different concentrations of metformin for 6 h. Mitochondrial respiratory chain activity was determined by using Seahorse XF96 Extracellular Flux Analyzers in Seahorse assay medium (0.55 mg/ml pyruvate in base medium, pH7.4). After determination of basal oxygen consumption rates, cells were sequentially treated with oligomycin A (1 μM), FCCP(1 μM), and rotenone (1 μM) along with antimycin A(1 μM). Viable cell numbers were counted and used to normalize the oxygen consumption rate.
Different metformin concentrations were used to determine the effects of metformin on mitochondrial activity of complex I (ab109903, abcam), complex II+III (ab109905, abcam), complex IV (ab109906, abcam), and complex V (ab109907, abcam) in in vitro assays following the manufacturer’s procedures.
Wang Y., An H., Liu T., Qin C., Sesaki H., Guo S., Radovick S., Hussain M., Maheshwari A., Wondisford F.E., O’Rourke B, & He L. (2019). Metformin Improves Mitochondrial Respiratory Activity through Activation of AMPK. Cell reports, 29(6), 1511-1523.e5.
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2

Mitochondrial Complex I Activity Assay

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Human neurons (5000 cells/well) were treated with medium alone (control cells) or GPE at three different concentrations (5, 50 and 500 μM) or with choline alphoscerate (500 μM) for 21 DIV. Then, chain respiratory complex I activity was determined by an in vitro ELISA (MitoTox Complex I OXPHOS activity assay, Abcam ab109903). Briefly, 50 μl of the diluted mitochondria to each well was added and incubated for 2 h at room temperature. After incubation time, 40 μl of PLs to each well was added and incubated for 45 min at room temperature. In fine, Complex I Activity solution was added for 1 h. The measure OD340 at 1‐min intervals for 2 h at 30°C was performed. The data were reported as the fold change from control cells set to 100%.
Zappelli E., Daniele S., Ceccarelli L., Vergassola M., Ragni L., Mangano G, & Martini C. (2022). α‐Glyceryl‐phosphoryl‐ethanolamine protects human hippocampal neurons from ageing‐induced cellular alterations. The European Journal of Neuroscience, 56(5), 4514-4528.
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3

Quantification of Energy Metabolites

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ATP, NADH and mitochondrial complex I were measured using the specific kit (ATP kit: Beyotime S0026, NADH kit: Beyotime S0175, mitochondrial complex I activity: Abcam ab109903) following the manufacturer's instructions.
Zhang X., Dong Y., Zhao M., Ding L., Yang X., Jing Y., Song Y., Chen S., Hu Q, & Ni Y. (2020). ITGB2-mediated metabolic switch in CAFs promotes OSCC proliferation by oxidation of NADH in mitochondrial oxidative phosphorylation system. Theranostics, 10(26), 12044-12059.
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4

Mitochondrial Respiratory Complex Analysis

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Cells were washed twice with phosphate-buffered saline and incubated for 30 min on ice in lysis buffer (68 mM sucrose, 200 mM mannitol, 50 mM potassium chloride, 1 mM ethylenediaminetetraacetic acid (EDTA), 1 mM EGTA, and 1 mM dithiothreitol with a protease inhibitor cocktail. The cells were then lysed using 45 passages through a 25G 5/8 needle and centrifuged at 1,500 g for 10 min. Cytosolic extracts were recovered after centrifugation at 13,000 g for 20 min. The pellet contained the mitochondria. The total protein concentration was determined using the BCA kit (Pierce, USA) according to the manufacturer’s instructions. The mitochondrial activity of complex I (ab109903, Abcam), complex II + III (b109905, Abcam), complex IV (ab109906, Abcam), and complex V (ab109907, Abcam) was determined using in vitro assays following the manufacturer’s procedures. The citrate synthase activity of mitochondrial extracts was measured using a Citrate Synthase Activity Assay Kit (ab119692, Abcam) according to the manufacturer’s instructions.
Ma Y., Chen G., Yi J., Li Q., Tan Z., Fan W., Luo X., He Z., Si Z, & Li J. (2022). IRX3 plays an important role in the pathogenesis of metabolic-associated fatty liver disease by regulating hepatic lipid metabolism. Frontiers in Endocrinology, 13, 895593.
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5

Metformin Effects on Hepatocyte Mitochondria

Check if the same lab product or an alternative is used in the 5 most similar protocols
Primary hepatocytes were seeded in an XF 96 well plate coated with 0.01% collagen type 1.36 h after seeding, cells were washed with PBS twice, and glucose production medium was added with different concentrations of metformin for 6 h. Mitochondrial respiratory chain activity was determined by using Seahorse XF96 Extracellular Flux Analyzers in Seahorse assay medium (0.55 mg/ml pyruvate in base medium, pH7.4). After determination of basal oxygen consumption rates, cells were sequentially treated with oligomycin A (1 μM), FCCP(1 μM), and rotenone (1 μM) along with antimycin A(1 μM). Viable cell numbers were counted and used to normalize the oxygen consumption rate.
Different metformin concentrations were used to determine the effects of metformin on mitochondrial activity of complex I (ab109903, abcam), complex II+III (ab109905, abcam), complex IV (ab109906, abcam), and complex V (ab109907, abcam) in in vitro assays following the manufacturer’s procedures.
Wang Y., An H., Liu T., Qin C., Sesaki H., Guo S., Radovick S., Hussain M., Maheshwari A., Wondisford F.E., O’Rourke B, & He L. (2019). Metformin Improves Mitochondrial Respiratory Activity through Activation of AMPK. Cell reports, 29(6), 1511-1523.e5.
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