mtTFA Antibody (18G102B2E11) - BSA Free

Simple Western: mtTFA Antibody (18G102B2E11) [NBP1-71648] - Simple Western lane view shows a specific band for mtTFA in 0.5 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

mtTFA-Antibody-18G102B2E11-Western-Blot-NBP1-71648-img0010.jpg

Immunocytochemistry/Immunofluorescence: mtTFA Antibody (18G102B2E11) [NBP1-71648] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-mtTFA Antibody (18G102B2E11) NBP1-71648 at 1 ug/ml overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

mtTFA-Antibody-18G102B2E11-Western-Blot-NBP1-71648-img0013.jpg

Immunohistochemistry-Paraffin: mtTFA Antibody (18G102B2E11) [NBP1-71648] - IHC staining of mtTFA on human testis using DAB with hematoxylin counterstain.

Immunocytochemistry/Immunofluorescence: mtTFA Antibody (18G102B2E11) [NBP1-71648] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-mtTFA Antibody (18G102B2E11) NBP1-71648 at 1 ug/ml overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunocytochemistry/Immunofluorescence: mtTFA Antibody (18G102B2E11) [NBP1-71648] - Immunostaining in HeLa cells. C1: DAPI staining 2 nuclei, C2: anti-mtTFA stain with Alexa Fluor 488 secondary ab (green), C3: Mitotracker Red staining mitochondria. Image courtesy of Elizabeth Wang.

Western Blot: mtTFA Antibody (18G102B2E11) [NBP1-71648] - Total protein from Hela and U937 were separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-mtTFA NBP1-71648 in blocking buffer and detected with an anti-mouse HRP secondary antibody using NovaLume chemiluminescence detection reagent (NPB2-61915).

Western Blot: mtTFA Antibody (18G102B2E11) [NBP1-71648] - Analysis of mtTFA expression in HeLa whole cell lysate.

Western Blot: mtTFA Antibody (18G102B2E11) [NBP1-71648] - MtDNA depletion & reversible impaired immune functions in THP-1 cells. A & B, Treatment with 50 ng/mL ethidium bromide (EtBr) for 8 weeks. A, MtDNA levels. B, LPS-induced TNF-alpha & IL-8 release. C-E, Transfection with 30 nmol/L negative or TFAM siRNA for 8 days. C, TFAM protein relative to beta -actin. D, MtDNA levels. E, LPS-induced TNF-alpha & IL-8 release. F-H, TFAM recovery 8 days after removal of TFAM siRNA. F, TFAM protein relative beta -actin. G, MtDNA levels. H, LPS-induced TNF-alpha & IL-8 release. All experiments are presented as means ± SD of 3 to 4 independent biological replicates. *P < .05, **P < .01, & ***P < .001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28629747), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: mtTFA Antibody (18G102B2E11) [NBP1-71648] - MtDNA depletion & impaired immune functions & subsequent recovery in THP-1 cells following transfection with TFAM siRNA. A-G, Transfection with TFAM siRNA. A, TFAM proteins levels relative to beta -actin during titration of TFAM siRNA, showing optimal knockdown of TFAM protein after transfection of THP-1 cells with 30 nmol/L siRNA for 8 days. B, Cell viability. C, Cell proliferation. D, The levels of the MT-CO1 & SDHA proteins relative to beta -actin. D, OCR for different aspects of mitochondrial respiration & respiratory profile. E, Phagocytosis of E coli. Recovery 8 days after removal of TFAM siRNA. F, Levels of the MT-CO1 & SDHA proteins relative to beta -actin. G, Oxygen consumption for different aspects of mitochondrial respiration. H, Bacterial phagocytosis. All experiments were carried out on 3 to 4 independent biological replicates & are presented as means ± SD. **P < .01 & ***P < .001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28629747), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

mtTFA (18G102B2E11) was detected in immersion fixed U-2 OS human osteosarcoma cell line using Mouse anti-mtTFA (18G102B2E11) Protein G Purified Monoclonal Antibody (Catalog # NBP1-71648) at 1.0 µg/mL overnight at 4C. Cells were stained using DyLight 488-conjugated Anti-Mouse IgG (H+L) Cross-Absorbed Secondary Antibody (green), and counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

mtTFA (18G102B2E11) was detected in immersion fixed U-2 OS human osteosarcoma cell line using Mouse anti-mtTFA (18G102B2E11) Protein G Purified Monoclonal Antibody conjugated to Alexa Fluor® 647 (Catalog # NBP1-71648AF647) (light blue) at 5 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

• Reactivity: Hu, Mu(-)
• Applications: WB, Simple Western, ICC/IF, IHC
• Clone: 18G102B2E11
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1 mg/ml

mtTFA Antibody (18G102B2E11) - BSA Free Summary

• Description: Novus Biologicals Mouse mtTFA Antibody (18G102B2E11) - BSA Free (NBP1-71648) is a monoclonal antibody validated for use in IHC, WB, ICC/IF and Simple Western. Anti-mtTFA Antibody: Cited in 17 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: Human mtTFA [Swiss-Prot# Q00059]
• Localization: Mitochondrion.
• Isotype: IgG2b Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: TFAM
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1-5 ug/ml
  • Immunohistochemistry 1:200
  • Immunohistochemistry-Paraffin 1:200
  • Simple Western 1:20
  • Western Blot 1:1000
• Application Notes: In Western blot, a band can be seen around 20 - 25 kDa. In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
  See Simple Western Antibody Database for Simple Western validation: Tested in HeLa lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:20, apparent MW was 33 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.
• Control:

  cIAP-2/HIAP-1 Overexpression Lysate

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein G purified

Alternate Names for mtTFA Antibody (18G102B2E11) - BSA Free

• mitochondrial transcription factor A
• MtTF1
• mtTFA
• TCF-6
• TCF6L1
• TCF6L2Mitochondrial transcription factor 1
• TCF6TCF6L3
• Transcription factor 6
• Transcription factor 6-like 2 (mitochondrial transcription factor)
• Transcription factor 6-like 2
• transcription factor A, mitochondrial

Background

mtTFA (transcription factor A, mitochondrial) is an essential protein that binds mitochondrial DNA (mtDNA) with or without sequence specificity to control mitochondrial transcription initiation and mtDNA copy number. mtTFA exists as monomer localized in mitochondrion matrix/nucleoid where it binds DNA and interacts with TFB1M/TFB2M. mtTFA associates with mitochondrial light strand promoter for controlling transcription and is essential for correct/efficient mtRNA polymerase promoter recognition. mtTFA is capable of unwinding DNA and bending mitochondrial light strand promoter DNA into a U-turn shape via its HMG boxes, and mtTFA is essential for controlling mtDNA levels as well as its organization/compaction. mtDNA is a multicopy, circular genome that is required for respiratory function and interference with mitochondrial genome sequence fidelity, transcription, or translation can cause ATP insufficiency and several pathologies including progressive external ophthalmoplegia, amyotrophic lateral sclerosis (ALS), Alpers' disease, Parkinson's disease, Alzheimer's disease, aging, cardiovascular disease, diabetes and cancer.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for mtTFA Antibody (NBP1-71648)(18)

Publications using NBP1-71648

• Xu W, Zhao L An Enzyme-Linked Immunosorbent Assay for the Detection of Mitochondrial DNA-Protein Cross-Links from Mammalian Cells DNA 2022-11-11 (Western Blot, Human)
• Zhang C, Shen S, Xu L et Al. LONP1 alleviates ageing-related renal fibrosis by maintaining mitochondrial homeostasis J Cell Mol Med 2024-09-11 [PMID: 39261902]
• Xu R, Huang L, Liu J et al. Remodeling of Mitochondrial Metabolism by a Mitochondria-Targeted RNAi Nanoplatform for Effective Cancer Therapy Small (Weinheim an der Bergstrasse, Germany) 2023-11-02 [PMID: 37919865]
• Xu W, Zhao L. An Enzyme-Linked Immunosorbent Assay for the Detection of Mitochondrial DNA-Protein Cross-Links from Mammalian Cell DNA (Basel) 2023-08-21 [PMID: 37601565] (Western Blot, Human)
• Ilamathi H, Benhammouda S, Lounas A et al. Contact sites between endoplasmic reticulum sheets and mitochondria regulate mitochondrial DNA replication and segregation iScience 2023-06-01 [PMID: 37534187] (ICC/IF, Human)
• Dey S, Catchpole T, Takacs A, Csaky KG. et Al. Investigating the effects of 7-ketocholesterol on retinal pigment epithelium bioenergetics FASEB J 2023-05-30 [PMID: 37249566]
• Saal KA, Shaib AH, Mougios N et al. Heat denaturation enables multicolor X10-STED microscopy Scientific reports 2023-04-01 [PMID: 37005431] (ICC/IF, Human)
• Vidali S, Feichtinger RG, Emberger M et al. Ageing is associated with a reduction in markers of mitochondrial energy metabolism in the human epidermis Experimental dermatology 2023-02-27 [PMID: 36851889] (Immunohistochemistry-Paraffin, Human)
• Limagne E, Nuttin L, Thibaudin M et al. MEK inhibition overcomes chemoimmunotherapy resistance by inducing CXCL10 in cancer cells Cancer cell 2022-01-17 [PMID: 35051357]
• Patrick S, Gowda P, Lathoria K Et al. Diminished YAP1 affects mitochondrial dynamics in IDH1 mutant glioma Journal of cell science 2021-10-15 [PMID: 34651186]
• SA Harper, JR Bassler, S Peramsetty, Y Yang, LM Roberts, D Drummer, RT Mankowski, C Leeuwenbur, K Ricart, RP Patel, MM Bamman, SD Anton, BC Jaeger, TW Buford Resveratrol and exercise combined to treat functional limitations in late life: A pilot randomized controlled trial Exp Gerontol, 2020-10-15;0(0):111111. 2020-10-15 [PMID: 33068691] (Simple Western, Human)
• Takishita Y, Yasuda H, Shimizu M et al. Formation of neutrophil extracellular traps in mitochondrial DNA-deficient cells J Clin Biochem Nutr 2020-01-01 [PMID: 32001952] (Western Blot, Human)
• Widdrington JD, Gomez-Duran A, Pyle A et al. Exposure of Monocytic Cells to Lipopolysaccharide Induces Coordinated Endotoxin Tolerance, Mitochondrial Biogenesis, Mitophagy, and Antioxidant Defenses. Front Immunol. 2018-09-27 [PMID: 30319656] (WB, Human)
• Dlaskova A, Engstova H, Spacek T et al. 3D super-resolution microscopy reflects mitochondrial cristae alternations and mtDNA nucleoid size and distribution Biochim Biochim Biophys Acta Bioenerg. 2018-05-01 [PMID: 29727614] (ICC/IF, Human)
  
  Details:
  Citation using the Alexa Fluor 647 form of this antibody.
• Huang C, Wang K, Yang Y et al. Gastrodia elata alleviates mutant huntingtin aggregation through mitochondrial function and biogenesis mediation Phytomedicine 2018-01-01 [PMID: 29433686]
• Widdrington JD, Gomez-Duran A, Steyn JS et al. Mitochondrial DNA depletion induces innate immune dysfunction rescued by IFN-gamma J. Allergy Clin. Immunol. 2017-06-17 [PMID: 28629747] (WB, Human)
• Sun Q, Zhong W, Zhang W, Zhou Z Defect of mitochondrial respiratory chain is a mechanism of ROS overproduction in a rat model of alcoholic liver disease: role of zinc deficiency. Am J Physiol Gastrointest Liver Physiol 2016-02-01 [PMID: 26585415]
• Choi J, Chandrasekaran K, Demarest TG et al. Brain diabetic neurodegeneration segregates with low intrinsic aerobic capacity. Annals of Clinical and Translational Neurology 2014-01-01 [PMID: 25356430]

Bioinformatics

• Gene Symbol: TFAM
• Entrez:
  • 7019(Human)
• Uniprot:
  • Q00059(Human)