Anti-TOMM20 antibody [EPR15581-39] - Mitochondrial Marker - Rat IgG2a (Chimeric)

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-TOMM20 antibody [EPR15581-39] - Mitochondrial Marker - Rat IgG2a (Chimeric) (AB289670)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human cervix adenocarcinoma epithelial cell line) cells labeling TOMM20 with ab289670 at 1/1000 dilution (Red) compared with a rat monoclonal IgG isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat F(ab)2 Anti-Rat IgG Fc (Alexa Fluor^® 488, ab150161) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TOMM20 antibody [EPR15581-39] - Mitochondrial Marker - Rat IgG2a (Chimeric) (AB289670)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labelling TOMM20 with ab289670 at 1/500 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human liver. The section was incubated with ab289670 for 30 mins at room temperature and followed by rabbit specific IgG antibody (ab102248) for 8 mins. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-TOMM20 antibody [EPR15581-39] - Mitochondrial Marker - Rat IgG2a (Chimeric) (AB289670)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labeling TOMM20 with ab289670 at 1/100 dilution, followed by Goat Anti-Rat IgG H&L (Alexa Fluor^® 488 (ab150157) secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HeLa cell line. The nuclear counterstain is DAPI (Blue). Tubuln is labeled using ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution (Red).

Secondary antibody only control : Used PBS instead of primary antibody, followed by Goat Anti-Rat IgG H&L (Alexa Fluor^® 488 (ab150157) secondary antibody at 1/1000 dilution.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-TOMM20 antibody [EPR15581-39] - Mitochondrial Marker - Rat IgG2a (Chimeric) (AB289670)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat small intestine tissue labelling TOMM20 with ab289670 at 1/50 dilution, followed by ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor^® 488) secondary antibody at 1/1000 dilution (Green). Positive staining on rat small intestine. The nuclear conterstain is DAPI (Blue).

Secondary antibody only control : Used PBS instead of primary antibody, followed by ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor^® 488) secondary antibody at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-TOMM20 antibody [EPR15581-39] - Mitochondrial Marker - Rat IgG2a (Chimeric) (AB289670)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse small intestine tissue labelling TOMM20 with ab289670 at 1/50 dilution, followed by ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor^® 488) secondary antibody at 1/1000 dilution (Green). Positive staining on mouse small intestine. The nuclear conterstain is DAPI (Blue).

Secondary antibody only control : Used PBS instead of primary antibody, followed by ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor^® 488) secondary antibody at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TOMM20 antibody [EPR15581-39] - Mitochondrial Marker - Rat IgG2a (Chimeric) (AB289670)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized Rat Primary Hippocampal Neurons DIV14 cells labelling ALDH2 with ab326874 at 1 ug/mL dilution (Green).

ab326874 staining ALDH2 in primary rat neurons/glia, DIV14 (prepared from E18 rat hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. SDHEP) cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal rabbit serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab326874 at 1µg/ml (shown in green), ab289670 - Anti-TOMM20 antibody [EPR15581-39] - Mitochondrial Marker - Rat IgG2a (Chimeric) (shown in pseudocolour magenta) and ab104224 - Ms Anti-NeuN antibody [1B7] - Neuronal Marker (shown in pseudocolour yellow). Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution, ab150167 - Goat Anti-Rat IgG H&L (Alexa Fluor® 647) preadsorbed at 1/1000 dilution and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue). Also suitable in cells fixed with 4% paraformaldehyde (10 min).Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

was used to counterstain tubulin at 1/ dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

• WB

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Western blot - Anti-TOMM20 antibody [EPR15581-39] - Mitochondrial Marker - Rat IgG2a (Chimeric) (AB289670)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-TOMM20 antibody [EPR15581-39] - Mitochondrial Marker - Rat IgG2a (Chimeric) (ab289670) at 1/1000 dilution

All lanes:

HeLa (human cervix adenocarcinoma epithelial cell line) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/5000 dilution

Predicted band size: 16 kDa

Observed band size: 16 kDa

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