Anti-mtTFA antibody [RM1035] - BSA and Azide free

• IHC

Supplier Data

Immunohistochemistry - Anti-mtTFA antibody [RM1035] - BSA and Azide free (AB307303)

This data was developed using ab307302, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labeling mtTFA with ab307302 at 1/1000 (0.54 ug/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on mitochondria in human lung carcinoma. The section was incubated with ab307302 for 30 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 a ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC

Supplier Data

Immunohistochemistry - Anti-mtTFA antibody [RM1035] - BSA and Azide free (AB307303)

This data was developed using ab307302, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling mtTFA with ab307302 at 1/1000 (0.54 ug/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on mitochondria in human colon. The section was incubated with ab307302 for 30 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 a ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-mtTFA antibody [RM1035] - BSA and Azide free (AB307303)

This data was developed using ab307302, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U-2 OS (human bone osteosarcoma epithelial cell) cells labeling mtTFA with ab307302 at 1/100 (5.4 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing mitochondrial staining in U-2 OS cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab197491 Anti-COX IV Mouse monoclonal antibody (Alexa Fluor® 647) - Mitochondrial Marker was used to counterstain tubulin at 1/200 dilution (2.5 ug/ml) 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 (2 ug/ml) dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-mtTFA antibody [RM1035] - BSA and Azide free (AB307303)

This data was developed using ab307302, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MCF7 (human breast adenocarcinoma epithelial cell) cells labelling mtTFA with ab307302 at 1/500 dilution (0.1ug) (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

Supplier Data

Immunoprecipitation - Anti-mtTFA antibody [RM1035] - BSA and Azide free (AB307303)

This data was developed using ab307302, the same antibody clone in a different buffer formulation.

mtTFA was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate 10µg with ab307302 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307302 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate 10µg
Lane 2 : ab307302 IP in MCF7 whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307302 in MCF7 whole cell lysate

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

Exposure time : 8 seconds

All lanes:

Immunoprecipitation - Anti-mtTFA antibody [RM1035] - Mitochondrial Marker (<a href='/en-us/products/primary-antibodies/mttfa-antibody-rm1035-mitochondrial-marker-ab307302'>ab307302</a>) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate 10 µg

Lane 2:

MCF7 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

Observed band size: 25 kDa

false

Exposure time: 8s

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-mtTFA antibody [RM1035] - BSA and Azide free (AB307303)

This data was developed using ab307302, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C2C12 (mouse myoblasts myoblast) cells labeling mtTFA with ab307302 at 1/100 (5.4 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing cytoplasmic staining in C2C12 cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 ug/ml) 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 (2 ug/ml) dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-mtTFA antibody [RM1035] - BSA and Azide free (AB307303)

This data was developed using ab307302, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized C2C12 (mouse myoblast) cells labelling mtTFA with ab307302 at 1/500 dilution (0.1ug) (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

Supplier Data

Immunoprecipitation - Anti-mtTFA antibody [RM1035] - BSA and Azide free (AB307303)

This data was developed using ab307302, the same antibody clone in a different buffer formulation.

mtTFA was immunoprecipitated from 0.35 mg C2C12 (mouse myoblast) whole cell lysate 10µg with ab307302 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307302 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : C2C12 (mouse myoblast) whole cell lysate 10µg
Lane 2 : ab307302 IP in C2C12 whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307302 in C2C12 whole cell lysate

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

Exposure time : 24 seconds

All lanes:

Immunoprecipitation - Anti-mtTFA antibody [RM1035] - Mitochondrial Marker (<a href='/en-us/products/primary-antibodies/mttfa-antibody-rm1035-mitochondrial-marker-ab307302'>ab307302</a>) at 1/1000 dilution

Lane 1:

C2C12 (mouse myoblast) whole cell lysate 10 µg

Lane 2:

C2C12 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/mttfa-antibody-rm1035-mitochondrial-marker-ab307302'>ab307302</a> in C2C12 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

Observed band size: 25 kDa

false

Exposure time: 24s

• WB

Supplier Data

Western blot - Anti-mtTFA antibody [RM1035] - BSA and Azide free (AB307303)

This data was developed using ab307302, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 70 seconds Exposure time :

All lanes:

Western blot - Anti-mtTFA antibody [RM1035] - Mitochondrial Marker (<a href='/en-us/products/primary-antibodies/mttfa-antibody-rm1035-mitochondrial-marker-ab307302'>ab307302</a>) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellar carcinoma epithelial cell) non-mitochondrial fraction 20 μg

Lane 2:

HepG2 mitochondria fraction 20 μg

Lane 3:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate 20 μg

Lane 4:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate 20 μg

Lane 5:

C2C12 (mouse myoblast) whole cell lysate 20 μg

Lane 6:

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

Lane 7:

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

Lane 8:

Mouse heart lysate 20 μg

Lane 9:

Rat heart lysate 20 μg

Secondary

All lanes:

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

Observed band size: 25 kDa

false

Exposure time: 70s