Overview

  • Product name

    Anti-mtTFA antibody [EPR12285]
    See all mtTFA primary antibodies
  • Description

    Rabbit monoclonal [EPR12285] to mtTFA
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WB, ICC/IF, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human mtTFA aa 50-150 (Cysteine residue). The exact sequence is proprietary.
    Database link: Q00059

  • Positive control

    • HeLa, A431, MCF7 and 293T cell lysates; Human kidney and prostate tissues; MCF7 cells.
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab176558 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P 1/300. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

For unpurified, use 1/50 - 1/100.

WB 1/10000 - 1/50000. Predicted molecular weight: 29 kDa.
ICC/IF 1/100 - 1/250.
IP 1/10 - 1/100.

Target

  • Function

    Binds to the mitochondrial light strand promoter and functions in mitochondrial transcription regulation. Required for accurate and efficient promoter recognition by the mitochondrial RNA polymerase. Promotes transcription initiation from the HSP1 and the light strand promoter by binding immediately upstream of transcriptional start sites. Is able to unwind and bend DNA. Required for maintenance of normal levels of mitochondrial DNA. May play a role in organizing and compacting mitochondrial DNA. target DNA. Interacts with TFB1M and TFB2M.
  • Sequence similarities

    Contains 2 HMG box DNA-binding domains.
  • Cellular localization

    Mitochondrion.
  • Information by UniProt
  • Database links

  • Alternative names

    • anscription factor 6-like 1 antibody
    • Mitochondrial transcription factor 1 antibody
    • mitochondrial transcription factor A antibody
    • MtTF1 antibody
    • mtTFA antibody
    • TCF 6 antibody
    • TCF-6 antibody
    • TCF6 antibody
    • TCF6L1 antibody
    • TCF6L2 antibody
    • TCF6L3 antibody
    • TFAM antibody
    • TFAM_HUMAN antibody
    • Transcription factor 6 antibody
    • Transcription factor 6 like 2 (mitochondrial transcription factor) antibody
    • Transcription factor 6 like 2 antibody
    • Transcription factor 6-like 2 antibody
    • transcription factor 6-like 3 antibody
    • Transcription factor A, mitochondrial antibody
    • Transcription factor A, mitochondrial antibody
    • Transcription factor A, mitochondrial precursor antibody
    see all

Images

  • All lanes : Anti-mtTFA antibody [EPR12285] (ab176558) at 1/10000 dilution (purified)

    Lane 1 : HeLa cell lysate
    Lane 2 : MCF7 cell lysate
    Lane 3 : HEK293 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

    Predicted band size: 29 kDa
    Observed band size: 24 kDa
    why is the actual band size different from the predicted?



    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Immunohistochemical staining of paraffin embedded human thyroid carcinoma with purified ab176558 at a working dilution of 1/300. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
  • Immunofluorescence staining of MCF7 cells with purified ab176558 at a working dilution of 1/250, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab176558 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

  • ab176558 (purified) at 1/40 immunoprecipitating mtTFA in 10 μg HEK293 (Lanes 1 and 2, observed at 24 kDa). Lane 3 - PBS. For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution. Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST

  • All lanes : Anti-mtTFA antibody [EPR12285] (ab176558) at 1/10000 dilution (unpurified)

    Lane 1 : HeLa cell lysate
    Lane 2 : A431 cell lysate
    Lane 3 : MCF7 cell lysate
    Lane 4 : 293T cell lysate

    Lysates/proteins at 10 µg per lane.

    Developed using the ECL technique.

    Predicted band size: 29 kDa

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human kidney tissue, labeling mtTFA using unpurified ab176558 at a 1/50 dilution.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human prostate tissue, labeling mtTFA using unpurified ab176558 at a 1/50 dilution.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunofluorescence analysis of MCF7 cells, labeling mtTFA using unpurified ab176558 at a 1/100 dilution.

  • Western blot analysis on Immunoprecipitation pellet from either 1) 293T cell lysate, or 2) 1xPBS (negative control), showing mtTFA, using unpurified ab176558 at 1/10 dilution and HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG.

References

This product has been referenced in:

  • Guo YX  et al. Effects of nutrient restriction and arginine treatment on oxidative stress in the ovarian tissue of ewes during the luteal phase. Theriogenology 113:127-136 (2018). WB ; Sheep . Read more (PubMed: 29501008) »
  • Huang Y  et al. Supplementing the maternal diet of rats with butyrate enhances mitochondrial biogenesis in the skeletal muscles of weaned offspring. Br J Nutr 117:12-20 (2017). Read more (PubMed: 28091351) »
See all 7 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Kidney
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Feb 29 2016

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