Overview

  • Product name
  • Description
    Mouse monoclonal to TOMM20
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, IHC-P, Flow Cyt, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Rat, Human
    Predicted to work with: Mouse
  • Immunogen

    Recombinant full length protein (GST-tag) corresponding to Human TOMM20 aa 1-145.
    Database link: Q15388

  • Positive control
    • WB: HeLa, PC-12 and NIH/3T3 cell lysates. IHC-P: Human small intestine. ICC/IF: HeLa cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab56783 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 Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 16 kDa.
IHC-P Use a concentration of 3 µg/ml.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IP Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 - 10 µg/ml.

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human small intestine tissue labelling TOMM20 with ab56783 at 3 µg/ml.

  • All lanes : Anti-TOMM20 antibody (ab56783)

    Lane 1 : HeLa cell lysate
    Lane 2 : PC-12 cell lysate
    Lane 3 : NIH/3T3 cell lysate

    Predicted band size: 16 kDa

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling TOMM20 with ab56783 at 10 µg/ml. Cells are fixed with 4% PFA and permeabilized on ice in PBS 0.1% Triton. Samples were incubated with primary antibody at 4oC overnight and fluorscein- conjugated secondary antibody at 4oC  for 1 hour.  

  • ab56783 staining TOMM20 in human keratinocytes by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with acetone and blocked with 1% BSA for 1 hour at 24°C. Samples were incubated with primary antibody (1/2000 in PBS + 1% BSA) for 24 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-mouse IgG polyclonal was used as the secondary antibody (1/5000).

    See Abreview

  • ab56783 staining TOMM20 in HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with PBS + 0.1% Triton X-100 and blocked with PBS+ 0.5% BSA + 0.2% fish skin gelatin for 1 hour at 25°C. Samples were incubated with primary antibody (1/2000 in PBS + 1% BSA) for 24 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-mouse IgG polyclonal was used as the secondary antibody (1/500).

    See Abreview

  • TOMM20 was immunoprecipitated using 0.5 mg HepG2 whole cell extract, 5 µg of Mouse monoclonal to TOMM20 (ab56783) and 50 µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10 min, HepG2 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10 min under agitation.
    Proteins were eluted by addition of 40 µl SDS loading buffer and incubated for 10 min at 70°C; 10 µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab56783.
    Secondary: Protein G-HRP at 1/500 dilution.
    Band: 14kDa: TOMM20. Non specific - 25kDa: We are unsure as to the identity of this extra band.

  • Overlay histogram showing HeLa cells stained with ab56783 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab56783, 1 μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was a mix of mouse IgG1 [ICIGG1], (ab91353, 2μg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4%PFA/permeabilized in 0.1% PBS-Tween used under the same conditions.

References

This product has been referenced in:
  • Deo P  et al. Outer membrane vesicles from Neisseria gonorrhoeae target PorB to mitochondria and induce apoptosis. PLoS Pathog 14:e1006945 (2018). Read more (PubMed: 29601598) »
  • Pan L  et al. miR-125a induces apoptosis, metabolism disorder and migrationimpairment in pancreatic cancer cells by targeting Mfn2-related mitochondrial fission. Int J Oncol 53:124-136 (2018). Read more (PubMed: 29749475) »
See all 49 Publications for this product

Customer reviews and Q&As

1-10 of 18 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Horse Cultured Cells (Equine tenocytes third passage)
Permeabilization
Yes - methanol
Specification
Equine tenocytes third passage
Blocking step
1%BSA, 10% horse serum, 0.1% Tween 20, 22.52mg/mL glycine in PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Methanol

Mr. David Bardell

Verified customer

Submitted Oct 09 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (GT1-7 hypothalamic neuronal)
Permeabilization
Yes - PBS + 0.3% Triton X-100
Specification
GT1-7 hypothalamic neuronal
Fixative
Paraformaldehyde

Dr. David Burk

Verified customer

Submitted Oct 04 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium citrate buffer
Permeabilization
No
Specification
Kidney
Blocking step
Fish skin gelatin as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.2% · Temperature: 21°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 06 2018

Application
Western blot
Sample
Mouse Cell lysate - whole cell (MEF cells and Hela cells)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
50 µg
Specification
MEF cells and Hela cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Aug 30 2018

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (muscle transverse section)
Permeabilization
Yes - triton 0.3%
Specification
muscle transverse section
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 29 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Inner medullary collecting duct (IMCD) cells)
Permeabilization
Yes - 0.5% Triton X-100
Specification
Inner medullary collecting duct (IMCD) cells
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 6% · Temperature: 22°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 28 2017

Application
Western blot
Sample
Mouse Tissue lysate - whole (Rat and mouse Hippocampus)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
20 µg
Specification
Rat and mouse Hippocampus
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Apr 03 2017

Application
Western blot
Sample
Mouse Cell lysate - whole cell (D2.0R, D2A1 murine mammary tumor cell lines)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
20 µg
Treatment
50 ´M Chloroquine for 48hrs
Specification
D2.0R, D2A1 murine mammary tumor cell lines
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Mar 27 2017

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Sample
Fruit fly (Drosophila melanogaster) Cell (heart tube myocyte)
Specification
heart tube myocyte
Permeabilization
Yes - 1% triton x-100
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jun 11 2013

Answer

Thank you for contacting us.



We searched information of TOMM20 protein in database, and according to the description on the database, TOMM20 is a mitochondrion outer membrane.(http://www.uniprot.org/uniprot/Q15388). As such this product should suit your needs.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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1-10 of 18 Abreviews or Q&A

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