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RecombinantRabMAb

Recombinant Anti-mtTFA antibody [RM1035] (ab307302)

Reviews (1) Submit a question
Western blot - Anti-mtTFA antibody [RM1035] (ab307302)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mtTFA antibody [RM1035] (ab307302)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mtTFA antibody [RM1035] (ab307302)
  • Immunocytochemistry/ Immunofluorescence - Anti-mtTFA antibody [RM1035] (ab307302)
  • Immunocytochemistry/ Immunofluorescence - Anti-mtTFA antibody [RM1035] (ab307302)
  • Flow Cytometry - Anti-mtTFA antibody [RM1035] (ab307302)
  • Flow Cytometry - Anti-mtTFA antibody [RM1035] (ab307302)
  • Immunoprecipitation - Anti-mtTFA antibody [RM1035] (ab307302)
  • Immunoprecipitation - Anti-mtTFA antibody [RM1035] (ab307302)
  • Anti-mtTFA antibody [RM1035] (ab307302)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit recombinant multiclonal [RM1035] to mtTFA
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-mtTFA antibody [RM1035]
    See all mtTFA primary antibodies

  • Description

    Rabbit recombinant multiclonal [RM1035] to mtTFA

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    This product was produced with the following immunogens:
    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HepG2 mitochondria / non-mitochondria fraction, MCF7, HEK-293, C2C12, NIH/3T3, and C6 whole cell lysates; mouse and rat heart tissue lysates. IHC-P: Human colon and lung carcinoma FFPE tissue sections. ICC/IF: U-2 OS and C2C12 cells. Flow Cyt: MCF7 and C2C12 cells. IP: MCF7 and C2C12 whole cell lysates.

  • General notes

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

    Recombinant multiclonals are a mixture of recombinant antibodies co-expressed from a library of heavy and light chains.

    Recombinant multiclonal antibodies offer the sensitivity of polyclonal antibodies by recognising multiple epitopes, along with consistency of a recombinant antibody.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab307302 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
WB (1)
1/1000. Detects a band of approximately 25 kDa.
IHC-P
1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/100.
Flow Cyt
1/500.
IP
1/30.
Notes
WB
1/1000. Detects a band of approximately 25 kDa.
IHC-P
1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/100.
Flow Cyt
1/500.
IP
1/30.

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

    see all

  • 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

  • Western blot - Anti-mtTFA antibody [RM1035] (ab307302)
    Western blot - Anti-mtTFA antibody [RM1035] (ab307302)
    All lanes : Anti-mtTFA antibody [RM1035] (ab307302) at 1/1000 dilution

    Lane 1 : HepG2 (human hepatocellar carcinoma epithelial cell) non-mitochondrial fraction
    Lane 2 : HepG2 mitochondria fraction
    Lane 3 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
    Lane 4 : HEK-293 (human embryonic kidney epithelial cell) whole cell lysate
    Lane 5 : C2C12 (mouse myoblast) whole cell lysate
    Lane 6 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
    Lane 7 : C6 (rat glial tumor glial cell) whole cell lysate
    Lane 8 : Mouse heart lysate
    Lane 9 : Rat heart lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

    Observed band size: 25 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST  

     Exposure time: 70 seconds

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mtTFA antibody [RM1035] (ab307302)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mtTFA antibody [RM1035] (ab307302)

    Immunohistochemical analysis of paraffin-embedded human colon tissue labeling mtTFA with AB307302 at 1/1000 (0.54 ug/ml) followed by ready to use Leica DS9800 (Bond® Polymer Refine Detection) kit. Positive staining on mitochondria in human colon. The section was incubated with ab307302 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is ready to use Leica DS9800 (Bond® Polymer Refine Detection) kit.

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mtTFA antibody [RM1035] (ab307302)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mtTFA antibody [RM1035] (ab307302)

    Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue labeling mtTFA with AB307302 at 1/1000 (0.54 µg/ml) followed by ready to use Leica DS9800 (Bond® Polymer Refine Detection) kit. Positive staining on mitochondria in human lung carcinoma. The section was incubated with ab307302 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is ready to use Leica DS9800 (Bond® Polymer Refine Detection) kit.

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

  • Immunocytochemistry/ Immunofluorescence - Anti-mtTFA antibody [RM1035] (ab307302)
    Immunocytochemistry/ Immunofluorescence - Anti-mtTFA antibody [RM1035] (ab307302)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U-2 OS (human bone osteosarcoma epithelial cell) cells labeling mtTFA with AB307302 at 1/100 (5.4 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 µg/ml) dilution (Green). Confocal image showing mitochondrial staining in U-2 OS cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab197491 Anti-COX IV Mouse monoclonal antibody (Alexa Fluor® 647) - Mitochondrial Marker was used to counterstain tubulin at 1/200 dilution (2.5 µg/ml) dilution (Red). The nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/ml) dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-mtTFA antibody [RM1035] (ab307302)
    Immunocytochemistry/ Immunofluorescence - Anti-mtTFA antibody [RM1035] (ab307302)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C2C12 (mouse myoblasts myoblast) cells labeling mtTFA with AB307302 at 1/100 (5.4 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 µg/ml) dilution (Green). Confocal image showing cytoplasmic staining in C2C12 cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/ml) dilution (Red). The nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/ml) dilution.

  • Flow Cytometry - Anti-mtTFA antibody [RM1035] (ab307302)
    Flow Cytometry - Anti-mtTFA antibody [RM1035] (ab307302)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MCF7 (human breast adenocarcinoma epithelial cell) cells labeling mtTFA with AB307302 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

  • Flow Cytometry - Anti-mtTFA antibody [RM1035] (ab307302)
    Flow Cytometry - Anti-mtTFA antibody [RM1035] (ab307302)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized C2C12 (mouse myoblast) cells labeling mtTFA with AB307302 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

  • Immunoprecipitation - Anti-mtTFA antibody [RM1035] (ab307302)
    Immunoprecipitation - Anti-mtTFA antibody [RM1035] (ab307302)

    mtTFA was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate 10µg with AB307302 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using AB307302 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

    Lane 1: MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate 10µg

    Lane 2: AB307302 IP in MCF7 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab307302 in MCF7 whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 8 seconds

     

  • Immunoprecipitation - Anti-mtTFA antibody [RM1035] (ab307302)
    Immunoprecipitation - Anti-mtTFA antibody [RM1035] (ab307302)

    mtTFA was immunoprecipitated from 0.35 mg C2C12 (mouse myoblast) whole cell lysate 10µg with AB307302 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using AB307302 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

    Lane 1: C2C12 (mouse myoblast) whole cell lysate 10µg

    Lane 2: AB307302 IP in C2C12 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab307302 in C2C12 whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 24 seconds

  • Anti-mtTFA antibody [RM1035] (ab307302)
    Anti-mtTFA antibody [RM1035] (ab307302)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (0)

Publishing research using ab307302? Please let us know so that we can cite the reference in this datasheet.

ab307302 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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Western blot (Dot Blot) abreview for Anti-mtTFA antibody [RM1035]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Purified protein (Purified Protein)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (Dot blot)
Loading amount
0.02 µg
Specification
Purified Protein
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Sean D Reardon

Verified customer

Submitted Mar 06 2023

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