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Anti tfam

Manufactured by Cell Signaling Technology
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Anti-TFAM is a primary antibody that recognizes Transcription Factor A, Mitochondrial (TFAM), a key regulator of mitochondrial DNA (mtDNA) transcription and replication. It can be used for the detection of TFAM protein in various cell and tissue samples using techniques such as Western blotting, immunohistochemistry, or immunocytochemistry.

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Lab products found in correlation

Anti β actin, by Merck Group (3 mentions) Anti nrf1, by Cell Signaling Technology (3 mentions) Irdye 680rd goat anti mouse, by LI COR (2 mentions) Cell lysis buffer, by Cell Signaling Technology (2 mentions) Anti α tubulin, by Merck Group (2 mentions) Hrp linked anti mouse igg, by Cell Signaling Technology (2 mentions) 4 to 20 polyacrylamide gels, by Bio-Rad (2 mentions) Odyssey fc analyzer, by LI COR (2 mentions) Anti β actin, by LI COR (2 mentions) Anti risp, by Abcam (2 mentions) Bovine serum albumin (bsa), by Merck Group (2 mentions) Anti ampkα, by Cell Signaling Technology (2 mentions) Trypsin, by Merck Group (1 mentions) Vitamin e, by Merck Group (1 mentions) Luperox, by Merck Group (1 mentions) Immun star hrp substrate kit, by Bio-Rad (1 mentions) Thiamine, by Merck Group (1 mentions) Pantethine, by Merck Group (1 mentions) Pantothenate, by Merck Group (1 mentions) Bodipy 581 591 c11, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Rabbit anti-TFAM (2 citations)

8 protocols using anti tfam

1

Antibody-based Mitochondrial Protein Analysis

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Monoclonal anti-actin antibody, Prussian Blue, pantothenate, pantethine, vitamin E, L-carnitine, thiamine, Luperox® and trypsin were purchased from Sigma Chemical Co. (St. Louis, MO). Anti-mitochondrial acyl carrier protein (mtACP), anti-NF-Y, anti-FOXN4 and anti-hnRNPA/B were purchased from Invitrogen/Molecular Probes (Eugene, OR). Anti-phospho-PGC1α was purchased from RD systems. NFS1 antibody and Omega 3 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Anti-PANK2, anti-PGC1α, complex 1 activity kit and PDH activity kit were purchased from Abcam (Cambridge, UK). Anti-TFAM was purchased from Cell Signaling. BODIPY® 581/591 C11 was purchased from Thermo-Fisher (Waltham, MA). A cocktail of protease inhibitors (complete cocktail) was purchased from Boehringer Mannheim (Indianapolis, IN). The Immun Star HRP substrate kit was from Bio-Rad Laboratories Inc. (Hercules, CA).
Álvarez-Córdoba M., Reche-López D., Cilleros-Holgado P., Talaverón-Rey M., Villalón-García I., Povea-Cabello S., Suárez-Rivero J.M., Suárez-Carrillo A., Munuera-Cabeza M., Piñero-Pérez R, & Sánchez-Alcázar J.A. (2022). Therapeutic approach with commercial supplements for pantothenate kinase-associated neurodegeneration with residual PANK2 expression levels. Orphanet Journal of Rare Diseases, 17, 311.
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2

Western Blot Analysis of Mitochondrial Proteins

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Lineage negative cells were purified following manufacturer's instruction (Stem Cell Technologies). Then, cells were lysed in cell lysis buffer (Cell Signaling), resolved on 4 to 20% polyacrylamide gels (BioRad) and transferred to nitrocellulose membranes. Membranes were blocked using 5% Milk for 1 hour and then incubated in the primary antibody overnight. Antibodies used were anti-RISP (Mitosciences; Catalog Number: ab14746. Used at a 1:500 dilution), anti-TFAM (a gift from Gerald Shadel, Yale University. Used at a 1:500 dilution), anti-β-actin (Sigma; Catalog Number: A2066. Used at 1:10,000 dilution) and anti-α-Tubulin (Sigma; Catalog Number: T9026. Used at a 1:2000 dilution). Membranes were washed 4 times with TBST and then incubated with secondary antibody. Secondary antibodies used were Anti-mouse IgG, HRP-Linked (Cell Signaling; Catalog Number: 7076S. 1:10,000 Dilution, used for anti-TFAM), Anti-rabbit IgG, HRP-Linked (Cell Signaling; Catalog Number: 7074S. 1:10,000 Dilution, used for anti-β-actin), and Goat Anti-Mouse IRDye 680RD (Li-cor; Catalog Number: C50721. 1:5,000 dilution. Used for anti-RISP and Anti-Tubulin). TFAM and actin blots were then treated wth ECL (Pierce) and developed using film. The RISP and tubulin blots were directly imaged using the Odyssey Fc Analyzer (Licor).
Ansó E., Weinberg S.E., Diebold L.P., Thompson B.J., Malinge S., Schumacker P.T., Liu X., Zhang Y., Shao Z., Steadman M., Marsh K.M., Xu J., Crispino J.D, & Chandel N.S. (2017). The mitochondrial respiratory chain is essential for haematopoietic stem cell function. Nature cell biology, 19(6), 614-625.
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3

Protein Expression Analysis in Cells

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Cells and tissues were lysed with RIPA buffer, and protein concentration was calculated by bicinchoninic acid assay. Proteins were isolated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride membranes. Membranes were blocked with 5% non-fat dry milk for 1 h at room temperature, and then incubated with primary antibodies including anti-AQP4 (Cell Signaling Technology, 59678S), anti-ASC (Cell Signaling Technology, 60824S), anti-AP2B1 (Santa Cruz, 74423), anti-CAV-1 (Santa Cruz, sc-53564), anti-NLRP3 (Life Science, mAG-20B-0014), anti-NRF1 (Abcam, ab175932), anti-Occludin (Proteintech, 66378), anti-p65 (Cell Signaling Technology, 8242), anti-p-p65 (Cell Signaling Technology, 3033S), anti-TFAM (Cell Signaling Technology, 8076S), and anti-β-actin (Sigma, A5316) overnight at 4°C. The binding of primary antibodies was visualized with goat anti-rabbit horseradish peroxidase (HRP)-conjugated secondary antibody (Jackson Laboratory, 115-035-033) or goat anti-mouse HRP-conjugated secondary antibody (Jackson Laboratory, 111-035-003).
Wang X., Chen G., Wan B., Dong Z., Xue Y., Luo Q., Wang D., Lu Y, & Zhu L. (2022). NRF1-mediated microglial activation triggers high-altitude cerebral edema. Journal of Molecular Cell Biology, 14(5), mjac036.
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4

Protein Expression Analysis by Western Blot

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Protein was extracted from cells cultured in 6-well plates using RIPA lysis buffer (Thermo Fisher Scientific, 89900) with protease inhibitors (Thermo Fisher Scientific, 78440). A total of 30-μg protein lysates was separated by 4%–12% NuPage Bis-Tris gels (Thermo Fisher Scientific, NP0336) and then transferred to polyvinylidene fluoride membranes (Bio-Rad, 1620264). The membranes were blocked with 5% BSA (Sigma-Aldrich, A7906–100G), probed with primary antibodies overnight at 4°C, and then incubated with the relevant secondary antibody for 1 hour at room temperature. Bands were visualized using ECL (Thermo Fisher Scientific, 34080) with Bio-Rad imaging system. The antibodies of anti-PARP (9532), anti-Rb (9309), anti-pRbSer807/811 (8516), anti-γH2AX (9718), anti-TFAM (8076), anti-AMPKα (5831), anti-pAMPKαThr172 (2535), anti-ERRα (13826), anti-VDAC (4661), anti-PHB1 (2426), anti-HSP60 (12165), anti-NRF2 (12721), anti-SDHA (11998), anti–β-actin (12620), anti–rabbit-IgG-HRP (7074), and anti–mouse-IgG-HRP (7076) were purchased from Cell Signaling Technology. The antibodies of anti-PGC1α (66369), anti-UQCRC1 (21705), anti-ATP5A1 (14676), anti-COXIV (11242), anti-NDUFB8 (14794), and anti-TUFM (26730) were purchased from Proteintech. Two or three independent experiments were performed.
Wei S., Yin D., Yu S., Lin X., Savani M.R., Du K., Ku Y., Wu D., Li S., Liu H., Tian M., Chen Y., Bowie M., Hariharan S., Waitkus M., Keir S.T., Sugarman E.T., Deek R.A., Labrie M., Khasraw M., Lu Y., Mills G.B., Herlyn M., Wu K., Liu L., Wei Z., Flaherty K.T., Abdullah K., Zhang G, & Ashley D.M. (2022). Antitumor Activity of a Mitochondrial-Targeted HSP90 Inhibitor in Gliomas. Clinical cancer research : an official journal of the American Association for Cancer Research, 28(10), 2180-2195.
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5

Western Blot Analysis of Mitochondrial Proteins

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Lineage negative cells were purified following manufacturer's instruction (Stem Cell Technologies). Then, cells were lysed in cell lysis buffer (Cell Signaling), resolved on 4 to 20% polyacrylamide gels (BioRad) and transferred to nitrocellulose membranes. Membranes were blocked using 5% Milk for 1 hour and then incubated in the primary antibody overnight. Antibodies used were anti-RISP (Mitosciences; Catalog Number: ab14746. Used at a 1:500 dilution), anti-TFAM (a gift from Gerald Shadel, Yale University. Used at a 1:500 dilution), anti-β-actin (Sigma; Catalog Number: A2066. Used at 1:10,000 dilution) and anti-α-Tubulin (Sigma; Catalog Number: T9026. Used at a 1:2000 dilution). Membranes were washed 4 times with TBST and then incubated with secondary antibody. Secondary antibodies used were Anti-mouse IgG, HRP-Linked (Cell Signaling; Catalog Number: 7076S. 1:10,000 Dilution, used for anti-TFAM), Anti-rabbit IgG, HRP-Linked (Cell Signaling; Catalog Number: 7074S. 1:10,000 Dilution, used for anti-β-actin), and Goat Anti-Mouse IRDye 680RD (Li-cor; Catalog Number: C50721. 1:5,000 dilution. Used for anti-RISP and Anti-Tubulin). TFAM and actin blots were then treated wth ECL (Pierce) and developed using film. The RISP and tubulin blots were directly imaged using the Odyssey Fc Analyzer (Licor).
Ansó E., Weinberg S.E., Diebold L.P., Thompson B.J., Malinge S., Schumacker P.T., Liu X., Zhang Y., Shao Z., Steadman M., Marsh K.M., Xu J., Crispino J.D, & Chandel N.S. (2017). The mitochondrial respiratory chain is essential for haematopoietic stem cell function. Nature cell biology, 19(6), 614-625.
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6

Immunofluorescence Analysis of Mitochondrial Regulators

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HLE and Huh7 cells following different treatments were seeded in confocal dishes (NEST, VA, USA) and maintained in DMEM for 24 h. The cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100, and then incubated with primary antibodies overnight at 4 ˚C. Antibodies were used as follows: anti-ZNF281 (1:50, Santa Cruz, CA, USA), anti-TFAM (1:50, Cell Signaling Technology Inc., Danvers, MA, USA), anti-NRF1 (1:50, Cell Signaling Technology Inc., Danvers, MA, USA), anti-PGC-1α (1:50, Millipore, Billerica, MA, USA), anti-SDHB (1:200, Abcam, Cambridge, MA, USA), and anti-MT-CO2 (1:200, Abcam, Cambridge, MA, USA). The fluorescent secondary antibodies used were Alexa fluor (m) 594 goat A or Alexa fluor (R) 488 goat A (1:200, Invitrogen, NY, USA). Counter-staining of the nuclei was performed using 4′, 6-diamidino-2-phenylindole (DAPI). Samples were observed under a Leica DM5000 B fluorescent microscope and Leica SP5 confocal microscope (Leica Microsystems, Wetzlar, Germany).
Zhao Q., Zhang C., Zhang X., Wang S., Guo T., Yin Y., Zhang H., Li Z., Si Y., Lu Y., Cheng S, & Ding W. (2023). ZNF281 inhibits mitochondrial biogenesis to facilitate metastasis of hepatocellular carcinoma. Cell Death Discovery, 9, 396.
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7

Melatonin Receptor Signaling Pathway Assay

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Pentanoic acid (HY-N6056), ROCK inhibitor-2 (HY-119937), S26131 (HY-122136) and 7-Desmethyl-agomelatine (HY-133113) were obtained from Med Chem Express (MCE, New Jersey, USA). SunP Gel G1 was purchased from SunP Biotech (Beijing, China). Bovine serum albumin (BSA) was purchased from Sigma-Aldrich. Anti-E-cadherin (ab1416), anti-MTNR1A (ab203038), anti-MTNR1B (ab203346), anti-tubulin (ab210797), Alexa 488 (ab150077) and Alexa 647 (ab150115) were purchased from Abcam (Cambridge, UK). Anti-COX4I1 (11242-1-AP) and anti-ATPB (17247-1-AP) were purchased from Proteintech (Hubei, China). Anti-TOM20 (#42406), anti-PGC1α (#2178), anti-NRF1 (#46743), and anti-TFAM (#8076) were obtained from Cell Signaling Technology (Massachusetts, USA). Anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH, TA309157) and all secondary antibodies were purchased from Zsbio (Beijing, China).
Zhang W., Zhao W., Li Q., Zhao D., Qu J., Yuan Z., Cheng Z., Zhu X., Zhuang X, & Zhang Z. (2021). 3D-printing magnesium–polycaprolactone loaded with melatonin inhibits the development of osteosarcoma by regulating cell-in-cell structures. Journal of Nanobiotechnology, 19, 263.
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8

Protein Analysis in Heart Tissues

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Heart tissues or cells were lysed using standard procedures, and the protein content was determined by BCA assay. Protein bands were detected using a Fluor Chem E imaging system (ProteinSimple, San Francisco, CA). Quantitation was performed using Image J software (National Institutes of Health, Bethesda, MD). β-actin was used as the loading control for total protein expression. The following primary antibodies were used: anti-acetyl lysine, Total OXPHOS Rodent WB Antibody, anti-peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α; all Abcam), anti-sirtuin 1 (Sirt1), anti-phospho-AMP-activated protein kinase (AMPK) α (Thr172), anti-AMPKα, anti-NRF1, anti-TFAM (all Cell Signaling Technology, Danvers, MA), and anti-β-actin (Santa Cruz Biotechnology, Santa Cruz, CA).
Tian J., Tang W., Xu M., Zhang C., Zhao P., Cao T., Shan X., Lu R, & Guo W. (2018). Shengmai San Alleviates Diabetic Cardiomyopathy Through Improvement of Mitochondrial Lipid Metabolic Disorder. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 50(5).
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