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.
  • General notes

    This product was changed from ascites to tissue culture supernatant on 22 May 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

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 at an assay dependent concentration. Predicted molecular weight: 16 kDa.
IHC-P Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

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 at an assay dependent concentration.

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.

    This image was generated using the ascites version of the product.

  • 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



    This image was generated using the ascites version of the product.

  • 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.  

    This image was generated using the ascites version of the product.

  • 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).

    This image was generated using the ascites version of the product.

    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).

    This image was generated using the ascites version of the product.

    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.

    This image was generated using the ascites version of the product.

  • 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.

    This image was generated using the ascites version of the product.

References

This product has been referenced in:

  • 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) »
  • Siekierska A  et al. Biallelic VARS variants cause developmental encephalopathy with microcephaly that is recapitulated in vars knockout zebrafish. Nat Commun 10:708 (2019). Read more (PubMed: 30755616) »
See all 68 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

Filter by Ratings

1-10 of 20 Abreviews

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Neural progenitor cell)
Permeabilization
Yes - 0.3% Triton X-100
Specification
Neural progenitor cell
Blocking step
(agent) for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 15 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa cells stably expressing Tomm20-GFP)
Permeabilization
Yes - 0.3% Triton X-100
Specification
HeLa cells stably expressing Tomm20-GFP
Blocking step
BSA as blocking agent for 15 minute(s) · Concentration: 1% · Temperature: 20°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 20 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Primary Fibroblast)
Permeabilization
Yes - Triton X-100 0.1%
Specification
Primary Fibroblast
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 14 2019

Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris)
Loading amount
20 µg
Specification
HeLa
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 Feb 06 2019

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

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

1-10 of 20 Abreviews

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up