mtTFA Antibody (18G102B2E11)

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.

Knockdown Validated: mtTFA Antibody (18G102B2E11) [NBP1-71648] - MtDNA depletion and reversible impaired immune functions in THP-1A cells. A and B, Treatment with 50A ng/mL ethidium bromide (EtBr) for 8A weeks. Transfection with 30A nmol/L negative or TFAM siRNA for 8A days. TFAM protein relative toI2-actin. D, MtDNA levels. E, LPS-induced TNF-I+-and IL-8 release. Image collected and cropped by Citeab from the following publication (Mitochondrial DNA depletion induces innate immune dysfunction rescued by IFN-g. J Allergy Clin Immunol (2017)) licensed under a CC-BY licence.

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.

Western Blot: mtTFA Antibody (18G102B2E11) [NBP1-71648] - Induction of mitochondrial biogenesis following exposure of THP-1 cells to LPS. THP-1 cells were incubated with LPS (100 ng/ml) for 0AE

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] - NIH3T3 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] - Alcohol feeding significantly decreased TFAM expression in rat liver. This image was submitted by a customer review.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, ICC/IF, IHC, IHC-P, KD, KO
• Clone: 18G102B2E11
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

mtTFA Antibody (18G102B2E11) Summary

• 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
  • Knockdown Validated
  • Knockout Validated
  • Simple Western 1:20
  • Western Blot 1:1000
• Application Notes: This mtTFA (18G102B2E11) antibody is useful for ICC/IF, IHC on paraffin-embedded sections and Western blot, where 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. Separated by Size-Wes, Sally Sue/Peggy Sue. Knockout validation (PMID: 32001952).
• Control:

  cIAP-2/HIAP-1 Overexpression Lysate

Reactivity Notes

Human and rat. Mouse reactivity reported in scientific literature (PMID: 28053692)

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: Tris-Glycine and 0.15M NaCl
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein G purified

Alternate Names for mtTFA Antibody (18G102B2E11)

• 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)(10)

Publications using NBP1-71648

• Patrick S, Gowda P, Lathoria K Et al. Diminished YAP1 affects mitochondrial dynamics in IDH1 mutant glioma Journal of cell science Oct 15 2021 [PMID: 34651186]
• Takishita Y, Yasuda H, Shimizu M et al. Formation of neutrophil extracellular traps in mitochondrial DNA-deficient cells J Clin Biochem Nutr Jan 1 2020 [PMID: 32001952] (WB, KO, Mouse)
• 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. Sep 27 2018 [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 May 1 [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 Jan 01 [PMID: 29433686] (Rat)
• 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 Jun 17 [PMID: 28629747] (WB, Human)
• Mastrocola R, Penna C et al. Pharmacological Inhibition of NLRP3 Inflammasome Attenuates Myocardial Ischemia/Reperfusion Injury by Activation of RISK and Mitochondrial Pathways. Oxid Med Cell Longev Jun 1 2017 [PMID: 28053692] (WB, Mouse)
• 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 Feb 1 [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 [PMID: 25356430]
• Morris EM, Meers GM, Booth FW et al. PGC-1alpha Overexpression Results in Increased Hepatic Fatty Acid Oxidation with Reduced Triacylglycerol Accumulation and Secretion Am J Physiol Gastrointest Liver Physiol 2012 Aug 16 [PMID: 22899824] (WB, Rat)

Reviews for mtTFA Antibody (NBP1-71648) (2)

Average Rating: 4

(Based on 2 reviews)

We have 2 reviews tested in 2 species: Mouse, Rat.

reviewed by: Prashant Rai ICC Mouse 08/27/2021

Summary

• Application: Immunocytochemistry
• Sample Tested: mice primary macrophages
• Species: Mouse
• Lot: A-8

Comments

• Comments: 0.1 % Triton X100 permeabilization

reviewed by: Verified Customer WB Rat 03/07/2018

Summary

• Application: Western Blot
• Sample Tested: Liver
• Species: Rat
• Lot: a1

Bioinformatics

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