HRP Anti-Glutaminase antibody [EP7212]

• WB

Lab

Western blot - HRP Anti-Glutaminase antibody [EP7212] (AB200408)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab200408 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

All lanes:

Western blot - HRP Anti-Glutaminase antibody [EP7212] (ab200408) at 1/5000 dilution

All lanes:

Kidney (Human) Tissue Lysate - fetal normal tissue at 10 µg

Predicted band size: 73 kDa

Observed band size: 73 kDa

true

Exposure time: 20min

• WB

Lab

Western blot - HRP Anti-Glutaminase antibody [EP7212] (AB200408)

ab200408 was shown to recognize Glutaminase in wild-type HAP1 cells as signal was lost at the expected MW in GLS (Glutaminase) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and GLS (Glutaminase) knockout samples were subjected to SDS-PAGE. ab200408 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor^® 680) loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/1000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

All lanes:

Western blot - HRP Anti-Glutaminase antibody [EP7212] (ab200408) at 1/5000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

GLS (Glutaminase) knockout HAP1 whole cell lysate at 20 µg

Predicted band size: 73 kDa

false

Exposure time: 12min