Anti-SLC25A12 antibody [EPR16294] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-SLC25A12 antibody [EPR16294] - BSA and Azide free (AB246329)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200201).

All lanes:

Immunoprecipitation - Anti-SLC25A12 antibody [EPR16294] (<a href='/en-us/products/primary-antibodies/slc25a12-antibody-epr16294-ab200201'>ab200201</a>)

Predicted band size: 74 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-SLC25A12 antibody [EPR16294] - BSA and Azide free (AB246329)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200201).

All lanes:

Immunoprecipitation - Anti-SLC25A12 antibody [EPR16294] (<a href='/en-us/products/primary-antibodies/slc25a12-antibody-epr16294-ab200201'>ab200201</a>)

Predicted band size: 74 kDa

false

• WB

Supplier Data

Western blot - Anti-SLC25A12 antibody [EPR16294] - BSA and Azide free (AB246329)

This data was developed using ab200201, the same antibody clone in a different buffer formulation.

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-SLC25A12 antibody [EPR16294] - BSA and Azide free (ab246329) at 1/5000 dilution

Lane 1:

Human fetal heart lysate at 20 µg

Lane 2:

Human skeletal muscle lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 74 kDa

Observed band size: 75 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-SLC25A12 antibody [EPR16294] - BSA and Azide free (AB246329)

This data was developed using ab200201, the same antibody clone in a different buffer formulation.

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-SLC25A12 antibody [EPR16294] - BSA and Azide free (ab246329) at 1/1000 dilution

Lane 1:

HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 10 µg

Lane 2:

A431 (Human epidermoid carcinoma) whole cell lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 74 kDa

Observed band size: 75 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-SLC25A12 antibody [EPR16294] - BSA and Azide free (AB246329)

This data was developed using ab200201, the same antibody clone in a different buffer formulation.

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:

Western blot - Anti-SLC25A12 antibody [EPR16294] - BSA and Azide free (ab246329)

Predicted band size: 74 kDa

Observed band size: 72 kDa

false

• WB

Supplier Data

Western blot - Anti-SLC25A12 antibody [EPR16294] - BSA and Azide free (AB246329)

This data was developed using ab200201, the same antibody clone in a different buffer formulation.

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-SLC25A12 antibody [EPR16294] - BSA and Azide free (ab246329) at 1/1000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Mouse heart lysate at 10 µg

Lane 3:

Mouse kidney lysate at 10 µg

Lane 4:

Rat brain lysate at 10 µg

Lane 5:

Rat heart lysate at 10 µg

Lane 6:

Rat kidney lysate at 10 µg

Lane 7:

C6 (Rat glial tumor cells) whole cell lysate at 10 µg

Lane 8:

RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 9:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 10 µg

Lane 10:

NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 74 kDa

Observed band size: 75 kDa

false

Exposure time: 15s