Mouse PRKAA1 knockout RAW 264.7 cell lysate

• WB

Lab

Western blot - Mouse PRKAA1 knockout RAW 264.7 cell lysate (AB280114)

Lane 1 : Wild-type RAW 264.7 cell lysate 20μg.

Lane 2 : PRKAA1 knockout RAW 264.7 cell lysate 20 μg.

Lane 3 : Mouse Liver cell lysate 20 μg.

Lane 4 : Neuro2A cell lysate 20 μg.

False colour image of Western blot : Anti-AMPK alpha 1 antibody [2B7] staining at 1/500 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution shown in red. In Western blot ab110036 was shown to bind specifically to AMPK alpha 1. A band was observed at 64 kDa in wild-type RAW 264.7 cell lysates with no signal observed at this size in PRKAA1 knockout cell line ab280055 (knockout cell lysate ab280114). To generate this image wild-type and PRKAA1 knockout RAW 264.7 cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

All lanes:

Western blot - Anti-AMPK alpha 1 antibody [2B7] (<a href='/ko/products/primary-antibodies/ampk-alpha-1-antibody-2b7-ab110036'>ab110036</a>) at 1/500 dilution

Lane 1:

Wild-type RAW 264.7 cell lysate at 20 µg

Lane 2:

PRKAA1 knockout RAW 264.7 cell lysate at 20 µg

Lane 2:

Western blot - Mouse PRKAA1 knockout RAW 264.7 cell line (<a href='/ko/products/cell-lines/mouse-prkaa1-knockout-raw-2647-cell-line-ab280055'>ab280055</a>)

Lane 3:

Mouse Liver cell lysate at 20 µg

Lane 4:

Neuro2A cell lysate at 20 µg

Predicted band size: 64 kDa

Observed band size: 64 kDa

false

• Sanger seq

Supplier Data

Sanger Sequencing - Mouse PRKAA1 knockout RAW 264.7 cell lysate (AB280114)

Mouse Prkaa1 KO in Raw264.7 Cells with 2 bp deletion in Exon 4

• WB

Lab

Western blot - Mouse PRKAA1 knockout RAW 264.7 cell lysate (AB280114)

Lane 1 : Wild-type RAW 264.7 cell lysate 20 μg

Lane 2 : PRKAA1 knockout RAW 264.7 cell lysate 20 μg

Lane 3 : Mouse Liver cell lysate 20 μg

Lane 4 : Neuro2A cell lysate 20 μg

False colour image of Western blot : Anti-AMPK alpha 1 antibody [Y365] staining at 1/1000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot ab32047 was shown to bind specifically to AMPK alpha 1. A band was observed at 64 kDa in wild-type RAW 264.7 cell lysates with no signal observed at this size in PRKAA1 knockout cell line ab280055 (knockout cell lysate ab280114). To generate this image wild-type and PRKAA1 knockout RAW 264.7 cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-AMPK alpha 1 antibody [Y365] (<a href='/ko/products/primary-antibodies/ampk-alpha-1-antibody-y365-ab32047'>ab32047</a>) at 1/1000 dilution

Lane 1:

Wild-type RAW 264.7 cell lysate at 20 µg

Lane 2:

Western blot - Mouse PRKAA1 knockout RAW 264.7 cell lysate (ab280114) at 20 µg

Lane 3:

Mouse Liver cell lysate at 20 µg

Lane 4:

Neuro2A cell lysate at 20 µg

Predicted band size: 64 kDa

Observed band size: 64 kDa

false