E2F4 Recombinant Rabbit Monoclonal Antibody [JE40-78]

Western blot analysis of E2F4 on different lysates with Rabbit anti-E2F4 antibody (HA721374) at 1/1,000 dilution.

Lane 1: K-562 cell lysate
Lane 2: A549 cell lysate
Lane 3: HEK-293 cell lysate
Lane 4: Jurkat cell lysate
Lane 5: HepG2 cell lysate
Lane 6: Raji cell lysate
Lane 7: HeLa cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 44 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 (HA721374) 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.

Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-E2F4 antibody (HA721374) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721374) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Flow cytometric analysis of K-562 cells labeling E2F4.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721374, 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).