AKT1 Recombinant Rabbit Monoclonal Antibody [PSH0-50]

Western blot analysis of AKT1 on different lysates with Rabbit anti-AKT1 antibody (HA721364) at 1/1,000 dilution.

Lane 1: Jurkat cell lysate
Lane 2: MCF7 cell lysate
Lane 3: HeLa cell lysate
Lane 4: NIH/3T3 cell lysate
Lane 5: C2C12 cell lysate
Lane 6: COS-1 cell lysate
Lane 7: PC-12 cell lysate
Lane 8: Mouse brain tissue lysate

Lysates/proteins at 30 µg/Lane.

Predicted band size: 56 kDa
Observed band size: 60 kDa

Exposure time: 3 minutes;

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 (HA721364) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry analysis of Jurkat cells labeling AKT1 with Rabbit anti-AKT1 antibody (HA721364) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-AKT1 antibody (HA721364) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Flow cytometric analysis of Jurkat cells labeling AKT1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721364, 1ug/ml) (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).