Overview

  • Product name
  • Description
    Mouse monoclonal to TOMM20
  • 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
  • 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

  • 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.  

  • 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:
  • Moretton A  et al. Selective mitochondrial DNA degradation following double-strand breaks. PLoS One 12:e0176795 (2017). WB . Read more (PubMed: 28453550) »
  • Shen Z  et al. PARK2-dependent mitophagy induced by acidic postconditioning protects against focal cerebral ischemia and extends the reperfusion window. Autophagy 13:473-485 (2017). Read more (PubMed: 28103118) »

See all 27 Publications for this product

Customer reviews and Q&As

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
Username

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
Username

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
Username

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
Username

Abcam user community

Verified customer

Submitted Jun 11 2013

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 ...

Read More
Application
Western blot
Sample
Mouse Cell lysate - other (Liver Mitochondrial fraction)
Loading amount
20 µg
Specification
Liver Mitochondrial fraction
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted May 15 2012

Thank you for contacting us. Anti-TOMM20 antibody (ab56783) has been protein G purified and is supplied in PBS at pH 7.2. I can therefore confirm that the antibody supplied does not contain any other carrier proteins or gelatine in the vial. I ...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human keratinocytes)
Specification
human keratinocytes
Fixative
Acetone
Permeabilization
No
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 24°C
Username

Abcam user community

Verified customer

Submitted May 27 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (keratinocytes)
Loading amount
80 µg
Specification
keratinocytes
Gel Running Conditions
Reduced Denaturing (15%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Apr 21 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Hippocampal culture)
Specification
Hippocampal culture
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Username

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Sep 01 2010

1-10 of 13 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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