Cofilin Recombinant Rabbit Monoclonal Antibody [PSH0-33]

Western blot analysis of Cofilin on different lysates with Rabbit anti-Cofilin antibody (HA721326) at 1/1,000 dilution.

Lane 1: Hela cell lysate (30 µg/Lane)
Lane 2: HEK-293 cell lysate (30 µg/Lane)
Lane 3: MCF-7 cell lysate (30 µg/Lane)
Lane 4: MDA-MB-468 cell lysate (24 µg/Lane)
Lane 5: K-562 cell lysate (16 µg/Lane)
Lane 6: SH-SY5Y cell lysate (22 µg/Lane)
Lane 7: HUVEC cell lysate (30 µg/Lane)
Lane 8: Jurkat cell lysate (30 µg/Lane)
Lane 9: A431 cell lysate (20 µg/Lane)
Lane 10: COS-1 cell lysate (30 µg/Lane)
Lane 11: VERO cell lysate (30 µg/Lane)
Lane 12: NIH/3T3 cell lysate (22 µg/Lane)
Lane 13: PC-12 cell lysate (30 µg/Lane)
Lane 14: Neuro-2a cell lysate (19 µg/Lane)

Predicted band size: 19 kDa
Observed band size: 19 kDa

Exposure time: 20 seconds;

15% 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 (HA721326) 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 1:200,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry analysis of NIH/3T3 cells labeling Cofilin with Rabbit anti-Cofilin antibody (HA721326) at 1/100 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-Cofilin antibody (HA721326) at 1/100 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 1/100 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 Hela cells labeling Cofilin.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721326, 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).

Flow cytometric analysis of NIH/3T3 cells labeling Cofilin.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721326, 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).