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Recombinant full length protein (GST-tag) corresponding to Human TOMM20 aa 1-145.
Database link: Q15388
Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. We would recommend antibody ab186734 as a replacement.
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.
|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.|
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human small intestine tissue labelling TOMM20 with ab56783 at 3 µg/ml.
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).
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).
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.
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