Phospho-AKT2 (S474) Recombinant Rabbit Monoclonal Antibody [PSH09-46] - BSA and Azide free

☑ Cell treatment (CT)

This data was developed using HA723108, the same antibody clone in a different buffer formulation.
Western blot analysis of Phospho-AKT2 (S474) on different lysates with Rabbit anti-Phospho-AKT2 (S474) antibody (HA723108) at 1/2,000 dilution.

Lane 1: MCF7 cell lysate
Lane 2: MCF7 treated with 50ng/mL Calyculin A for 45 minutes cell lysate
Lane 3: HEK-293 cell lysate
Lane 4: HEK-293 treated with 50μM LY294002 for 6 hours cell lysate
Lane 5: NIH/3T3 cell lysate
Lane 6: NIH/3T3 treated with 100ng/mL PDGF for 1 hour cell lysate
Lane 7: C6 cell lysate
Lane 8: C6 treated with 100ng/mL Calyculin A for 1 hour cell lysate

Lysates/proteins at 30 µg/Lane.

Predicted band size: 56 kDa
Observed band size: 56 kDa

Exposure time: Lane 1-6: 3 minutes; Lane 7-8: 12 seconds; ECL: K1802;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723108) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

☑ Cell treatment (CT)

This data was developed using HA723108, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of MCF7 cells (left) / MCF7 cells treated with 50ng/mL Calyculin A for 45 minutes (right) labeling Phospho-AKT2 (S474).

Cells were fixed and permeabilized. Then stained with the primary antibody (HA723108, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

☑ Cell treatment (CT)

This data was developed using HA723108, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of C6 cells (left) / C6 cells treated with 100ng/mL Calyculin A for 1 hours (right) labeling Phospho-AKT2 (S474).

Cells were fixed and permeabilized. Then stained with the primary antibody (HA723108, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).