Anti-Glycogen synthase 1/GYS1 antibody [EP817Y]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Glycogen synthase 1/GYS1 antibody [EP817Y] (AB40810)

Flow cytometry overlay histogram showing wild-type HeLa (green line) and GYS1 knockout HeLa stained with ab40810 (magenta line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab40810) (1x 10⁶ in 100μl at 0.04 μg/ml (1/15800)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C.

Isotype control antibody was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control in HeLa WT cells (black line) and HeLa-GYS1 KO cells (grey line), used at the same concentration and conditions as the primary antibody.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

This antibody gave a positive signal in wild-type HeLa fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

• WB

Lab

Western blot - Anti-Glycogen synthase 1/GYS1 antibody [EP817Y] (AB40810)

ab40810 Recombinant Anti-Glycogen synthase 1/GYS1 antibody [EP817Y] was shown to specifically react with Glycogen synthase 1/GYS1 in HeLa wild-type cells (ab255928). Loss of signal was observed when knockout cell line ab265388 (knockout cell lysate ab257462) was used. Wild-type and Glycogen synthase 1/GYS1 knockout samples were subjected to SDS-PAGE. ab40810 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Glycogen synthase 1/GYS1 antibody [EP817Y] (ab40810) at 1/10000 dilution

Lane 1:

Wild-type HeLa lysate at 20 µg

Lane 2:

Glycogen synthase 1/GYS1 knockout HeLa lysate at 20 µg

Lane 2:

Western blot - Human GYS1 (Glycogen synthase 1) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-gys1-glycogen-synthase-1-knockout-hela-cell-line-ab265388'>ab265388</a>)

Lane 3:

HEK-293 lysate at 20 µg

Lane 4:

Daudi lysate at 20 µg

Predicted band size: 84 kDa

Observed band size: 80 kDa

false

• WB

Unknown

Western blot - Anti-Glycogen synthase 1/GYS1 antibody [EP817Y] (AB40810)

All lanes:

Western blot - Anti-Glycogen synthase 1/GYS1 antibody [EP817Y] (ab40810) at 1/50000 dilution

All lanes:

HeLa cell lysate at 10 µg

Predicted band size: 84 kDa

Observed band size: 85 kDa

false