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Tom40

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TOM40 is a protein that is a core component of the translocase of the outer membrane (TOM) complex in mitochondria. The TOM complex is responsible for the import of nuclear-encoded proteins into the mitochondrial outer membrane.

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Lab products found in correlation

Mtch2, by Proteintech (3 mentions) Pdhe1 a, by Abcam (3 mentions) Tom20, by Santa Cruz Biotechnology (3 mentions) Beta actin, by Boster Bio (3 mentions) Mmp 9, by Cell Signaling Technology (3 mentions) Glial fibrillary acidic protein (gfap), by Merck Group (3 mentions) Protease and phosphatase inhibitor, by Thermo Fisher Scientific (3 mentions) Total oxphos rodent antibody cocktail, by Abcam (2 mentions) Phospho s6 ser240 244, by Cell Signaling Technology (2 mentions) 4 hne, by Abcam (2 mentions) Bcl2 associated x (bax), by Merck Group (2 mentions) Phospho 4e bp1 thr37 46, by Cell Signaling Technology (2 mentions) Phospho akt thr308, by Cell Signaling Technology (2 mentions) Cleaved caspase 3, by Cell Signaling Technology (2 mentions) N cadherin, by Cell Signaling Technology (2 mentions) Vimentin, by Cell Signaling Technology (2 mentions) Phospho akt ser473, by Cell Signaling Technology (2 mentions) Bcl 2, by Santa Cruz Biotechnology (2 mentions) Total oxphos human wb antibody cocktail, by Abcam (1 mentions) Hsp90, by Santa Cruz Biotechnology (1 mentions)

6 protocols using tom40

1

Western Blot Analysis of OXPHOS and DNA Damage

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Ten to 20 µg of protein per lane was separated by standard SDS–PAGE and transferred onto PVDF membranes. After blocking in blocking solution (BS; 100 mM Tris/HCl (pH 8.0), 450 mM NaCl, 5% dry milk, and 0.05% Tween-20), the membranes were incubated with antibodies against Total OXPHOS Human WB Antibody Cocktail (1:5000 in BS, Abcam), TOM40 (1:5000), p53 (1:5000 in BS, both from Santa Cruz Biotechnology, Dallas, TX, USA), and phospho-Ser139 Histone H2AX (γH2AX) (1:5000 in BS, Millipore/Merck KGaA, Darmstadt, Germany). Equal loading of whole cell lysates was verified by the detection of HSP90 (1:5000 in BS, Santa Cruz). Peroxidase-conjugated anti-mouse IgG (H + L) and anti-rabbit IgG (H + L) secondary antibodies (1:10,000; Abcam) were used and detection of specific signals was done with SuperSignal West Pico Chemiluminescent Substrate (Pierce/Thermo Scientific). Densitometric quantification was done with ImageJ (National Institutes of Health, Bethesda, MD, USA). Full immunoblot images are depicted in supplementary Fig. S4.
Marx C., Sonnemann J., Maddocks O.D., Marx-Blümel L., Beyer M., Hoelzer D., Thierbach R., Maletzki C., Linnebacher M., Heinzel T, & Krämer O.H. (2022). Global metabolic alterations in colorectal cancer cells during irinotecan-induced DNA replication stress. Cancer & Metabolism, 10, 10.
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2

Western Blot Analysis of Mitochondrial and Autophagy Markers

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Extracts for western blot were homogenized in RIPA buffer. In brief, proteins were separated by electrophoresis and then transferred to polyvinylidene difluoride membranes, which were processed with primary antibodies. Next, we incubated these membranes in the appropriate diluted secondary antibody at room temperature for 1 h. The follow primary antibodies were used: p62 (1:1000; Abcam, catalog no. ab56416), LC3 (1:1000; Cell Signaling Technology, catalog no. 2775s), translocase of outer mitochondrial membrane 40 homolog (TOM40; 1:1000; Abcam, catalog no. ab51884), COXIV (1:1000; Cell Signaling Technology, catalog no. 11967), manganese superoxide dismutase (MnSOD; 1:1000; Cell Signaling Technology, catalog no. 13141), β-actin (1:1000; Cell Signaling Technology, catalog no. 3700), and NeuN (1:1000; Abcam, catalog no. ab104224).
Lin C., Li N., Chang H., shen Y., Li Z., wei W., Chen H., Lu H., Ji J, & Liu N. (2020). Dual effects of thyroid hormone on neurons and neurogenesis in traumatic brain injury. Cell Death & Disease, 11(8), 671.
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3

Immunoblotting Analysis of Cardiac Protein Markers

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LV samples from DCM and control hearts were homogenized in a Laemmli buffer (253mM Tris/HCL pH 6.8, 8% SDS, 40% glycerin, 200mM DTT, 0.4% bromophenol blue). Proteins were quantified using the BCA Assay (Thermo Scientific Pierce Protein Biology, Schwerte, Germany). Equal amounts of total proteins were separated on SDS-polyacrylamide gels and transferred to a nitrocellulose membrane. The membranes were immunoblotted overnight with the following primary antibodies: AMPK (1:2000, Cell Signaling, USA), p-AMPK (1:2000, Thr172, Cell Signaling, USA), PGC-1α (1:1000 Abcam, UK), TOM40 (1:1000, Abcam, UK), TIM 23 (1:5000, BD, USA), Sirt3 (1:1000, Cell Signaling, USA), SOD2 (1:1000, Santa Cruz, USA), catalase (1:1000, Cell Signaling, USA), NFκBp65 (1:200, Santa Cruz, USA) and FOXO1 (1:1000, Cell Signaling, USA). Equal sample loading was confirmed by analysis of actin (1:1000, Santa Cruz, USA), HSP60 (1:1000, Cell Signaling, USA) or Ponceau S staining. Immunoreactive proteins were detected using ECL Plus (GE Healthcare, Buckinghamshire, UK) and quantified with ImageLab (version 5.2.1 build 11, Bio-Rad Laboratories (USA)).
Barcena M.L., Pozdniakova S., Haritonow N., Breiter P., Kühl A.A., Milting H., Baczko I., Ladilov Y, & Regitz-Zagrosek V. (2020). Dilated cardiomyopathy impairs mitochondrial biogenesis and promotes inflammation in an age- and sex-dependent manner. Aging (Albany NY), 12(23), 24117-24133.
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4

Quantifying Protein Expression Profiles

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Total cells were lysed with 2% SDS solution plus protease and phosphatase inhibitors (Thermo Scientific). Protein concentrations were tested by BCA kit and equivalent proteins were loaded into SDS-PAGE. Following western blots were performed according to standard procedures. The primary antibodies were list as follow: MTCH2 (Proteintech, Cat#16888-1-AP), Beta actin (Boster Biological Technology, Cat#BM0627), PDHE1-A (Abcam, Cat#ab110334), GFAP (Millipore, Cat#G3893), Tom20 (Santa Cruz Biotechnology, Cat#sc-136211), Tom40 (Abcam, Cat#ab185543), MMP-9 (Cell Signaling Technology, Cat#3852), N-cadherin (Cell Signaling Technology, Cat#4061), Vimentin (Cell Signaling Technology, Cat#5741), 4-HNE (Abcam, Cat#ab48506), BcL-2 (Santa Cruz Biotechnology, Cat#sc-7382), Bax (Millipore, Cat#06-499), phospho-AKT (Thr308) (Cell Signaling Technology, Cat#2965), phospho-AKT (Ser473) (Cell Signaling Technology, Cat#4060), AKT (Cell Signaling Technology, Cat#9272), phospho-S6 (Ser240/244) (Cell Signaling Technology, Cat#2215), S6 (Cell Signaling Technology, Cat#2217), phospho-4EBP1 (Thr37/46) (Cell Signaling Technology, Cat#9459), total OXPHOs rodent antibody cocktail (Abcam, Cat#110413), phospho-Cofilin1 (Ser3) (Cell Signaling Technology, Cat#3313), Cofilin1 (Cell Signaling Technology, Cat#5175), PARP (Abclonal, Cat#A0942) and cleaved caspase 3 (Cell Signaling Technology, Cat#9661).
Yuan Q., Yang W., Zhang S., Li T., Zuo M., Zhou X., Li J., Li M., Xia X., Chen M, & Liu Y. (2021). Inhibition of mitochondrial carrier homolog 2 (MTCH2) suppresses tumor invasion and enhances sensitivity to temozolomide in malignant glioma. Molecular Medicine, 27, 7.
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5

Protein Expression Profiling Protocol

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Total cells were lysed with 2% SDS solution plus protease and phosphatase inhibitors (Thermo Scienti c). Protein concentrations were tested by BCA kit and equivalent proteins were loaded into SDS-PAGE. Following western blots were performed according to standard procedures. The primary antibodies were list as follow: MTCH2 (Proteintech, Cat#16888-1-AP), Beta actin (Boster Biological Technology, Cat#BM0627), PDHE1-A (Abcam, Cat#ab110334), GFAP (Millipore, Cat#G3893), Tom20 (Santa Cruz Biotechnology, Cat#sc-136211), Tom40 (Abcam, Cat#ab185543), MMP-9 (Cell Signaling Technology,
Yuan Q., Yang W., Zhang S., Li T., Zuo M., Zhou X., Li J., Mao L., Xia X., Chen M., & Liu Y. (2020). Inhibition of Mitochondrial Carrier Homolog 2 (MTCH2) Suppresses Tumor Invasion and Enhances Sensitivity to Temozolomide in Malignant Glioma.
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6

Comprehensive Western Blot Analysis

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Total cells were lysed with 2% SDS solution plus protease and phosphatase inhibitors (Thermo Scienti c). Protein concentrations were tested by BCA kit and equivalent proteins were loaded into SDS-PAGE. Following western blots were performed according to standard procedures. The primary antibodies were list as follow: MTCH2 (Proteintech, Cat#16888-1-AP), Beta actin (Boster Biological Technology, Cat#BM0627), PDHE1-A (Abcam, Cat#ab110334), GFAP (Millipore, Cat#G3893), Tom20 (Santa Cruz Biotechnology, Cat#sc-136211), Tom40 (Abcam, Cat#ab185543), MMP-9 (Cell Signaling Technology, Cat#3852), N-cadherin (Cell Signaling Technology, Cat#4061), Vimentin (Cell Signaling Technology, Cat#5741), 4-HNE (Abcam, Cat#ab48506), BcL-2 (Santa Cruz Biotechnology, Cat#sc-7382), Bax (Millipore, Cat#06-499), phospho-AKT (Thr308) (Cell Signaling Technology, Cat#2965), phospho-AKT (Ser473) (Cell Signaling Technology, Cat#4060), AKT (Cell Signaling Technology, Cat#9272), phospho-S6 (Ser240/244) (Cell Signaling Technology, Cat#2215), S6 (Cell Signaling Technology, Cat#2217), phospho-4EBP1 (Thr37/46) (Cell Signaling Technology, Cat#9459), total OXPHOs rodent antibody cocktail (Abcam, Cat#110413), phospho-Co lin1 (Ser3) (Cell Signaling Technology, Cat#3313), Co lin1 (Cell Signaling Technology, Cat#5175), PARP (Abclonal, Cat#A0942) and cleaved caspase 3 (Cell Signaling Technology, Cat#9661).
Yuan Q., Yang W., Zhang S., Li T., Zuo M., Zhou X., Li J., Mao L., Xia X., Chen M., & Liu Y. (2020). Inhibition of Mitochondrial Carrier Homolog 2 (MTCH2) Suppresses Tumor Invasion and Enhances Sensitivity to Temozolomide in Malignant Glioma .
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