Anti-mtTFA antibody [EPR23548-120] - Mitochondrial Marker

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-mtTFA antibody [EPR23548-120] - Mitochondrial Marker (AB252432)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized C2C12 cells labelling mtTFA with ab252432 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic staining in C2C12 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 ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 2 ug/ml dilution.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-mtTFA antibody [EPR23548-120] - Mitochondrial Marker (AB252432)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PC-12 cells labelling mtTFA with ab252432 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic staining in PC-12 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 ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 2 ug/ml dilution.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-mtTFA antibody [EPR23548-120] - Mitochondrial Marker (AB252432)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized C2C12 (Mouse myoblasts myoblast) cells labelling mtTFA with ab252432 at 1/600 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, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-mtTFA antibody [EPR23548-120] - Mitochondrial Marker (AB252432)

mtTFA was immunoprecipitated from 0.35 mg PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate using ab252432 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab252432 at 1/500 dilution, followed by VeriBlot for IP Detection Reagent (HRP)(ab131366) at 1/1000 dilution.

Lane 1 : PC-12 whole cell lysate (Input)
Lane 2 : ab252432 IP in PC-12 cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab252432 in PC-12 whole cell lysate

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

Exposure time : 32 seconds

All lanes:

Immunoprecipitation - Anti-mtTFA antibody [EPR23548-120] - Mitochondrial Marker (ab252432)

Predicted band size: 29 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-mtTFA antibody [EPR23548-120] - Mitochondrial Marker (AB252432)

mtTFA was immunoprecipitated from 0.35 mg C2C12 (mouse myoblasts myoblast) whole cell lysate using ab252432 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab252432 at 1/500 dilution, followed by VeriBlot for IP Detection Reagent (HRP)(ab131366) at 1/1000 dilution.

Lane 1 : C2C12 whole cell lysate (Input)
Lane 2 : ab252432 IP in C2C12 cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab252432 in C2C12 whole cell lysate

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

Exposure time : 32 seconds

All lanes:

Immunoprecipitation - Anti-mtTFA antibody [EPR23548-120] - Mitochondrial Marker (ab252432)

Predicted band size: 29 kDa

false

• WB

Lab

Western blot - Anti-mtTFA antibody [EPR23548-120] - Mitochondrial Marker (AB252432)

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

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-mtTFA antibody [EPR23548-120] - Mitochondrial Marker (ab252432) at 1/1000 dilution

Lane 1:

Mouse cerebellum tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

Rat cerebellum tissue lysate at 20 µg

Lane 5:

Rat heart 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 1/50000 dilution

Predicted band size: 29 kDa

Observed band size: 25 kDa

false

• WB

Lab

Western blot - Anti-mtTFA antibody [EPR23548-120] - Mitochondrial Marker (AB252432)

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 25375380)

Exposure times : Lane 1 : 3 minutes; Lanes 2-5 : 70 seconds.

All lanes:

Western blot - Anti-mtTFA antibody [EPR23548-120] - Mitochondrial Marker (ab252432) at 1/1000 dilution

Lane 1:

C2C12 (mouse myoblasts myoblast) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 4:

PC-12 (rat adrenal gland pheochromocytoma ) whole cell lysate at 20 µg

Lane 5:

NIH/3T3 (mouse embryonic fibroblast) whole cell 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 1/50000 dilution

Predicted band size: 29 kDa

Observed band size: 25 kDa

false