Overview

  • Product name

    Anti-TOMM20 antibody - Mitochondrial Marker
    See all TOMM20 primary antibodies
  • Description

    Rabbit polyclonal to TOMM20 - Mitochondrial Marker
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IP, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Orangutan
  • Immunogen

    Synthetic peptide corresponding to Human TOMM20 aa 100 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab88207)

  • Positive control

    • This antibody gave a positive signal in HepG2 whole cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab78547 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
ICC/IF Use a concentration of 1 µg/ml.
IP Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 16 kDa (predicted molecular weight: 16 kDa).
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • Function

    Central component of the receptor complex responsible for the recognition and translocation of cytosolically synthesized mitochondrial preproteins. Together with TOM22 functions as the transit peptide receptor at the surface of the mitochondrion outer membrane and facilitates the movement of preproteins into the TOM40 translocation pore.
  • Sequence similarities

    Belongs to the Tom20 family.
  • Cellular localization

    Mitochondrion outer membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • KIAA0016 antibody
    • MAS20 antibody
    • MGC117367 antibody
    • Mitochondrial 20 kDa outer membrane protein antibody
    • Mitochondrial import receptor subunit TOM20 homolog antibody
    • MOM19 antibody
    • Outer mitochondrial membrane receptor Tom20 antibody
    • TOM20 antibody
    • TOM20_HUMAN antibody
    • TOMM20 antibody
    • Translocase of outer mitochondrial membrane 20 homolog (yeast) antibody
    • Translocase of outer mitochondrial membrane 20 homolog type II antibody
    see all

Images

  • ICC/IF image of ab78547 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab78547, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa, Hek293, MCF-7 cells at 1µg/ml, and in 100% Methanol fixed (5 min) HepG2, HeLa, Hek293, MCF-7 cells at 1µg/ml.
  • Anti-TOMM20 antibody - Mitochondrial Marker (ab78547) at 1 µg/ml + HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 16 kDa
    Observed band size: 16 kDa
    Additional bands at: 70 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 30 seconds
  • IHC image of TOMM20 staining in Human Liver carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab78547, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  • TOMM20 was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Rabbit polyclonal to TOMM20 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, HepG2 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab78547.
    Secondary: Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.
    Band: 16kDa; TOMM20.

References

This product has been referenced in:

  • Liu H  et al. Mitophagy protects SH-SY5Y neuroblastoma cells against the TNFa-induced inflammatory injury: Involvement of microRNA-145 and Bnip3. Biomed Pharmacother 109:957-968 (2019). Read more (PubMed: 30551550) »
  • Zhang YY  et al. CKD autophagy activation and skeletal muscle atrophy-a preliminary study of mitophagy and inflammation. Eur J Clin Nutr N/A:N/A (2019). Read more (PubMed: 30607007) »
See all 31 Publications for this product

Customer reviews and Q&As

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1-6 of 6 Abreviews

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Hippocampal neurons)
Permeabilization
Yes - TritonX-100
Specification
Hippocampal neurons
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 17 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (retina)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citric buffer pH 6
Permeabilization
Yes - triton x-100
Specification
retina
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 03 2019

Application
Western blot
Sample
Human Cell lysate - other (human fibroblast mitochondria)
Gel Running Conditions
Reduced Denaturing
Loading amount
100 µg
Specification
human fibroblast mitochondria
Blocking step
BSA as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jul 27 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (mouse embryonic fibroblast)
Specification
mouse embryonic fibroblast
Fixative
Formaldehyde
Permeabilization
Yes - 0.5% Triton X
Blocking step
BSA and image-iTFX (invitrogen) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 15 2013

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Formaldehyde
Permeabilization
Yes - 0.1% Triton X-100/PBS
Blocking step
0.5% BSA, 0.2% fish skin gelatin/PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted May 28 2010

Abreviews
Application
Western blot
Sample
Xenopus laevis Cell lysate - whole cell (meiotic egg extract)
Loading amount
25 µg
Specification
meiotic egg extract
Gel Running Conditions
Reduced Denaturing (15%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted May 21 2010

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