Ab192423

Manufactured by Abcam

Ab192423 is a laboratory equipment product. It is a scientific instrument used for specific applications in research and development. The core function of this product is to perform a particular set of tasks related to scientific analysis and experimentation. No further details can be provided in an unbiased and factual manner without potential extrapolation.

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Lab products found in correlation

3 protocols using ab192423

Mitochondrial Protein Expression Analysis

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Muscle biopsies and cybrid cells were lysed in cell lysis buffer (Beyotime, China). 30‐50 mg protein were run on a 8‐12% sodium dodecyl sulfate‐polyacrylamide gel (SDS–PAGE) at 100 V, 2‐3 h. Proteins were transferred to PVDF (Immobilon‐P PVDF‐Membrane, Millipore, Burlington, MA, USA) for 1‐3 h at 100 V, 4°C. After blocking with 5% dried skimmed milk, the membrane was incubated with various primary antibodies. Primary antibodies were as follows: total OXPHOS antibody cocktail (ab110413, Abcam), anti‐CO4 (CST 4805s, Cell Signaling Technology), anti‐GAPDH (10494‐1‐AP, Proteintech), anti‐ND5 antibody(ab92624, Abcam), anti‐ATP6 antibody (ab192423, Abcam), anti‐ND1 antibody (ab181848, Abcam), Anti‐CYB antibody (ab81215, Abcam), Anti‐VDAC1/Porin antibody (ab158995, Abcam), Anti‐β actin antibody (TA‐09, Zhongshan Jinqiao).

Ji K., Wang W., Lin Y., Xu X., Liu F., Wang D., Zhao Y, & Yan C. (2020). Mitochondrial encephalopathy Due to a Novel Pathogenic Mitochondrial tRNAGln m.4349C>T Variant. Annals of Clinical and Translational Neurology, 7(6), 980-991.

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Mitochondrial Protein Expression Analysis in B16 Cells

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B16 cells were transfected with Myg1 SiRNA using Dharmafect (SMARTpool:ON-TARGETplus Myg1 siRNA, L-055338-01-0005). After 24 h, the cells were trypsinized and lysed using NP40 lysis buffer supplemented with protease inhibitor cocktail. For detection of mitochondrial protein, 40 μg of cell lysate was resolved on 10% SDS-PAGE gel. Antibody was prepared in 2% skimmed milk solution with following dilutions MT-CO1(Abcam, ab90668, 1:500), MT-ND5(Abcam, ab92624, 1:500), MT-ATP6 (Abcam, ab192423, 1:100), MT-CYB(Abcam, ab81215, 1:100), Myg1 (Abcam, ab122493, 1:2000), Tubulin (Abcam, Ab21058, 1:5000). It was detected using HRP conjugated anti rabbit secondary antibody (1:10 000) and detected through Enhanced Chemiluminescence using ChemiDoc (Syngene).

Grover R., Burse S.A., Shankrit S., Aggarwal A., Kirty K., Narta K., Srivastav R., Ray A.K., Malik G., Vats A., Motiani R.K., Thukral L., Roy S.S., Bhattacharya S., Sharma R., Natarajan K., Mukerji M., Pandey R., Gokhale R.S, & Natarajan V.T. (2019). Myg1 exonuclease couples the nuclear and mitochondrial translational programs through RNA processing. Nucleic Acids Research, 47(11), 5852-5866.

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Comprehensive Mitochondrial Protein Analysis

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For Western blot analysis, the following antibodies were used in a 1:1000 dilution: anti-β (ab14730, Abcam), anti-γ (PA5-29975, ThermoFisher), anti-b (ab117991, Abcam), anti-OSCP (ab110276, Abcam), anti-f (ab200715, Abcam), anti-g (ab126181, Abcam), anti-e (ab122241, Abcam), anti-c (ab181243, Abcam), anti-a (ab192423, Abcam), anti-citrate synthase (ab96600, Abcam), anti-vinculin (V4505, Sigma), anti-prohibitin (MS-261-P1, NeoMarkers), anti-GAPDH (2118, Cell Signaling), anti-CyPD (ab110324, Abcam), anti-ANT2 (14671, Cell Signaling), anti-ANT3 (PA5-35113, ThermoFischer), anti-OXPHOS (ab110411, Abcam), anti-HKII (sc130358, Santa Cruz), anti-UQCRC1 (sc65238, Santa Cruz), anti-Grim19 (sc271013, Santa Cruz), and anti-SDHA (sc166947, Santa Cruz). Statistical comparison of data was assessed with the two-sided Student’s t-test.

Carrer A., Tommasin L., Šileikytė J., Ciscato F., Filadi R., Urbani A., Forte M., Rasola A., Szabò I., Carraro M, & Bernardi P. (2021). Defining the molecular mechanisms of the mitochondrial permeability transition through genetic manipulation of F-ATP synthase. Nature Communications, 12, 4835.

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