Anti-TOM70 antibody [EPR26576-162] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TOM70 antibody [EPR26576-162] - BSA and Azide free (AB290002)

This data was developed using ab289977, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon tissue labelling TOM70 with ab289977 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human colon. The section was incubated with ab289977 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 10 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TOM70 antibody [EPR26576-162] - BSA and Azide free (AB290002)

This data was developed using ab289977, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labelling TOM70 with ab289977 at 1/2000 dilution followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human breast cancer. The section was incubated with ab289977 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 10 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TOM70 antibody [EPR26576-162] - BSA and Azide free (AB290002)

This data was developed using ab289977, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labelling TOM70 with ab289977 at 1/2000 dilution followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human cerebrum. The section was incubated with ab289977 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 10 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TOM70 antibody [EPR26576-162] - BSA and Azide free (AB290002)

This data was developed using ab289977, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human esophagus tissue labelling TOM70 with ab289977 at 1/2000 dilution followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human esophagus. The section was incubated with ab289977 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 10 mins

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-TOM70 antibody [EPR26576-162] - BSA and Azide free (AB290002)

This data was developed using ab289977, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol, permeabilized U-2 OS (human bone osteosarcoma epithelial cell) cells labelling TOM70 with ab289977 at 1/500 dilution (0.1μg) (Red) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TOM70 antibody [EPR26576-162] - BSA and Azide free (AB290002)

This data was developed using ab289977, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U-2 OS cells labeling TOM70 with ab289977 at 1/100 dilution, followed by ab150081 Goat Anti-Rbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing mitochondrial staining in U-2 OS cell line. ab33985 Anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used at 1/1000 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TOM70 antibody [EPR26576-162] - BSA and Azide free (AB290002)

This data was developed using ab289977, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 cells labeling TOM70 with ab289977 at 1/100 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 μg/ml) dilution (Green). Confocal image showing cytoplasmic staining in NIH/3T3 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TOM70 antibody [EPR26576-162] - BSA and Azide free (AB290002)

This data was developed using ab289977, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C6 cells labeling TOM70 with ab289977 at 1/100 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in C6 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-TOM70 antibody [EPR26576-162] - BSA and Azide free (AB290002)

This data was developed using ab289977, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol, permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling TOM70 with ab289977 at 1/500 dilution (0.1μg) (Red) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-TOM70 antibody [EPR26576-162] - BSA and Azide free (AB290002)

This data was developed using ab289977, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized C6 (rat glial tumor glial cell) cells labelling TOM70 with ab289977 at 1/500 dilution (0.1μg) (Red) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IP

Lab

Immunoprecipitation - Anti-TOM70 antibody [EPR26576-162] - BSA and Azide free (AB290002)

This data was developed using ab289977, the same antibody clone in a different buffer formulation.

TOM70 was immunoprecipitated from NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab289977 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab289977 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 μg

Lane 2 : ab289977 IP in NIH/3T3 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab289977 in NIH/3T3 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3.25 seconds

All lanes:

Immunoprecipitation - Anti-TOM70 antibody [EPR26576-162] (<a href='/en-us/products/primary-antibodies/tom70-antibody-epr26576-162-ab289977'>ab289977</a>)

Predicted band size: 67 kDa

Observed band size: 70 kDa

false

• IP

Lab

Immunoprecipitation - Anti-TOM70 antibody [EPR26576-162] - BSA and Azide free (AB290002)

This data was developed using ab289977, the same antibody clone in a different buffer formulation.

TOM70 was immunoprecipitated from C6 (rat glial tumor glial cell) whole cell lysate with ab289977 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab289977 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : C6 (rat glial tumor glial cell) whole cell lysate 10 μg

Lane 2 : ab289977 IP in C6 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab289977 in C6 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3.25 seconds

All lanes:

Immunoprecipitation - Anti-TOM70 antibody [EPR26576-162] (<a href='/en-us/products/primary-antibodies/tom70-antibody-epr26576-162-ab289977'>ab289977</a>)

Predicted band size: 67 kDa

Observed band size: 70 kDa

false

• WB

Supplier Data

Western blot - Anti-TOM70 antibody [EPR26576-162] - BSA and Azide free (AB290002)

False colour image of Western blot : Anti-TOM70 antibody [EPR26576-162] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab289977 was shown to bind specifically to TOM70. A band was observed at 72 kDa in wild-type HeLa cell lysates with no signal observed at this size in TOMM70 knockout cell line ab265396 (knockout cell lysate ab258732). To generate this image, wild-type and TOMM70 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution. This data was developed using ab289977, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-TOM70 antibody [EPR26576-162] (<a href='/en-us/products/primary-antibodies/tom70-antibody-epr26576-162-ab289977'>ab289977</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human TOMM70A (TOM70) knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-tomm70a-tom70-knockout-hela-cell-lysate-ab258732'>ab258732</a>) at 20 µg

Lane 3:

U-2 OS cell lysate at 20 µg

Lane 4:

Mouse Brain cell lysate at 20 µg

Lane 5:

Rat Brain cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 67 kDa

Observed band size: 72 kDa

false

• WB

Lab

Western blot - Anti-TOM70 antibody [EPR26576-162] - BSA and Azide free (AB290002)

This data was developed using 289977, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

The expression profile/molecular weight observed is consistent with that described in the literature (PMID : 31907385, PMID : 11956321).

Exposure time : 3.25 seconds

All lanes:

Western blot - Anti-TOM70 antibody [EPR26576-162] (<a href='/en-us/products/primary-antibodies/tom70-antibody-epr26576-162-ab289977'>ab289977</a>) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 67 kDa

Observed band size: 70 kDa

false

• WB

Lab

Western blot - Anti-TOM70 antibody [EPR26576-162] - BSA and Azide free (AB290002)

This data was developed using 289977, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

The expression profile/molecular weight observed is consistent with that described in the literature (PMID : 32356556)

Low expression : Breast (human protein atlas : https : //www.proteinatlas.org/ENSG00000154174-TOMM70)

Exposure time : 15 seconds

All lanes:

Western blot - Anti-TOM70 antibody [EPR26576-162] (<a href='/en-us/products/primary-antibodies/tom70-antibody-epr26576-162-ab289977'>ab289977</a>) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 2:

Mouse brain tissue lysate at 20 µg

Lane 3:

Mouse liver tissue lysate at 20 µg

Lane 4:

Mouse breast tissue lysate at 20 µg

Lane 5:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 6:

Rat brain tissue lysate at 20 µg

Lane 7:

Rat liver tissue lysate at 20 µg

Lane 8:

Rat breast tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 67 kDa

Observed band size: 70 kDa

false

• IP

Lab

Immunoprecipitation - Anti-TOM70 antibody [EPR26576-162] - BSA and Azide free (AB290002)

This data was developed using ab289977, the same antibody clone in a different buffer formulation.

TOM70 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg with ab289977 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab289977 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg Lane 2 : ab289977 IP in HeLa whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab289977 in HeLa whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-TOM70 antibody [EPR26576-162] (<a href='/en-us/products/primary-antibodies/tom70-antibody-epr26576-162-ab289977'>ab289977</a>) at 1/30 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 10 µg

Lane 2:

<a href='/en-us/products/primary-antibodies/tom70-antibody-epr26576-162-ab289977'>ab289977</a> IP in HeLa whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/tom70-antibody-epr26576-162-ab289977'>ab289977</a> in HeLa whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Predicted band size: 67 kDa

Observed band size: 67 kDa

false

Exposure time: 3.25s