Atp5o

Manufactured by Abcam

Sourced in Germany

ATP5O, also known as ATP synthase subunit O, is a core component of the mitochondrial ATP synthase complex. It plays a crucial role in the production of cellular energy in the form of ATP through the process of oxidative phosphorylation. The ATP5O protein is essential for the proper assembly and function of the ATP synthase complex, which is responsible for the final step of energy production in the mitochondria.

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3 protocols using atp5o

Western Blot Analysis of Mitochondrial and Epigenetic Proteins

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Western blot was performed as described^{2 (link)} using the following antibodies: TFAM (7495S, Cell Signaling Technology; sc-23588, Santa Cruz Technology), PHB2 (14085S, Cell Signaling Technology), ATP5A1 (sc-136178, Santa Cruz Technology), ATP5B (sc-16690, Santa Cruz Technology), ATP5D (ab97491, Abcam), ATP5O (ab110276, Abcam, clone 4C11C10D12), 4EBP1 (9644S, Cell Signaling Technology), phos-4EBP1 (Thr37/46) (2855S, Cell Signaling Technology), p70S6K (2708S, Cell Signaling Technology), phos-S6K (Ser235/236) (2211S, Cell Signaling Technology), GATA1 (ab47490, Abcam), GATA2 (ab22849, Abcam), TSC1 (6935, Cell Signaling Technology), TSC2 (4308, Cell Signaling Technology), H3K9ac (9649, Cell Signaling Technology), H3K27ac (8173, Cell Signaling Technology), H3K56ac (4243, Cell Signaling Technology), H4K5ac (ab51997, Abcam), H3K4me3 (9571, Cell Signaling Technology), H3K9me3 (13969, Cell Signaling Technology), H3K27me3 (9733, Cell Signaling Technology), H3K36me3 (4909, Cell Signaling Technology), H3 (4499, Cell Signaling Technology), ACTB (MAB1501, Millipore, clone C4), and GAPDH (sc-26778, Santa Cruz Biotechnology). Densitometry quantification was performed using ImageJ software. All antibodies were used at 1:1,000 dilutions except phos-4EBP1 (1:500), phos-S6K (1:500) and ACTB (1:2,500).

Liu X., Zhang Y., Ni M., Cao H., Signer R.A., Li D., Li M., Gu Z., Hu Z., Dickerson K.E., Weinberg S.E., Chandel N.S., DeBerardinis R.J., Zhou F., Shao Z, & Xu J. (2017). Regulation of Mitochondrial Biogenesis in Erythropoiesis by mTORC1-Mediated Protein Translation. Nature cell biology, 19(6), 626-638.

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Western Blot Analysis of Mitochondrial Proteins

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The sample for the western blotting was prepared by scraping the cells in PBS. The protein extraction was done using RIPA lysis buffer. The amount of protein was determined using Bradford reagent and spectrophotometer for equal loading of a gel. The samples were prepared in Laemmli loading buffer. 10–20% precast Tricine gel (Invitrogen) was used (particularly for MIC13 as it is a small protein). The proteins were transferred onto nitrocellulose membrane and probed for various antibodies. Anti-HRP secondary antibodies were used. We used following antibodies for immunoblotting, MIC13 (Pineda, Berlin, polyclonal antibody raised in rabbit using following peptide CKAREYSKEGWEYVKARTK), MIC27/APOOL (Atlas Antibodies, HPA000612), MIC26/APOO (Thermo-Fisher, MA5-15493) MIC60/Mitofilin (Pineda, Berlin, polyclonal antibody raised in rabbit against human IMMT using the peptide CTDHPEIGEGKPTPALSEEAS), MIC10/MINOS1 (Pineda, Berlin, polyclonal antibody raised in rabbits against CQHDFQAPYLLHGKYVK), MIC25/CHCHD6 (cell signaling), VDAC (abcam), β-tubulin (Cell signaling), MIC19/CHCHD3 (abcam), ATP5L (proteintech), ATP5O (abcam), COXIV (abcam), NDUFB4 (abcam), UQCRC2 (abcam), MTND1 (abcam), TOM20 (Proteintech), TIM23 (BD biosciences), human TAZ1 (gift from Steve Claypool). Recording and visualization of chemiluminescent signals were done using VILBER LOURMAT Fusion SL (Peqlab).

Anand R., Strecker V., Urbach J., Wittig I, & Reichert A.S. (2016). Mic13 Is Essential for Formation of Crista Junctions in Mammalian Cells. PLoS ONE, 11(8), e0160258.

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Western Blot Analysis of Mitochondrial and Epigenetic Proteins

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