Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: SLC25A12 knockout HAP1 cell lysate (20 µg)
Lane 3: Human skeletal muscle lysate (20 µg)
Lane 4: A431 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab200201 observed at 72 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab200201 was shown to recognize SLC25A12 when SLC25A12 knockout samples were used, along with additional cross-reactive bands. Wild-type and SLC25A12 knockout samples were subjected to SDS-PAGE. ab200201 and ab8245 (loading control to GAPDH) were diluted to 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed ab216773 andGoat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes : Anti-SLC25A12 antibody [EPR16294] (ab200201) at 1/5000 dilution

Lane 1 : Human fetal heart lysate
Lane 2 : Human skeletal muscle lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 75 kDa
Observed band size: 75 kDa


Exposure time: 10 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

All lanes : Anti-SLC25A12 antibody [EPR16294] (ab200201) at 1/1000 dilution

Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 2 : A431 (Human epidermoid carcinoma) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 75 kDa
Observed band size: 75 kDa


Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

All lanes : Anti-SLC25A12 antibody [EPR16294] (ab200201) at 1/1000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Mouse kidney lysate
Lane 4 : Rat brain lysate
Lane 5 : Rat heart lysate
Lane 6 : Rat kidney lysate
Lane 7 : C6 (Rat glial tumor cells) whole cell lysate
Lane 8 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
Lane 9 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 10 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 75 kDa
Observed band size: 75 kDa


Exposure time: 15 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

SLC25A12 was immunoprecipitated from 1mg of Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate with ab200201 at 1/40 dilution.

Western blot was performed from the immunoprecipitate using ab200201 at 1/1000 dilution.

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1: Jurkat whole cell lysate 10 µg (Input).

Lane 2: ab200201 IP in Jurkat whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab200201 in Jurkat whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 second.

SLC25A12 was immunoprecipitated from 1mg of Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate with ab200201 at 1/40 dilution.

Western blot was performed from the immunoprecipitate using ab200201 at 1/1000 dilution (Panel A) or ab192244 at 1/1000 dilution (Panel B).

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1: Jurkat whole cell lysate 10 µg (Input).

Lane 2: ab200201 IP in Jurkat whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab200201 in Jurkat whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time (Panel A and Panel B): 30 second.