Anti-YWHAE antibody [RP23040165]

• WB

Supplier Data

Western blot - Anti-YWHAE antibody [RP23040165] (AB308051)

Western blot analysis was performed on Tissue extracts of Mouse Brain (Lane 1), Rat Brain (Lane 2) and Membrane extract of Jurkat (Lane 3), PC-12 (Lane 4), SW480 (Lane 5), NIH/3T3 (Lane 6), Hep G2 (Lane 7), HeLa (Lane 8) and T-47D (Lane 9). The blots were probed with ab308051 at 1-2 µg/mL dilution and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1/2500 dilution. A 29 kDa band corresponding to YWHAE was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using a 4-12% Bis-Tris gel, electrophoresis system and pre-stained protein standard. Resolved proteins were then transferred onto a nitrocellulose membrane. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed (ECL).

All lanes:

Western blot - Anti-YWHAE antibody [RP23040165] (ab308051) at 1 µg/mL

Lane 1:

Mouse brain at 30 µg

Lane 2:

Rat Brain at 30 µg

Lane 3:

Jurkat membrane extract at 30 µg

Lane 4:

PC-12 Membrane extract at 30 µg

Lane 5:

SW480 Membrane extract at 30 µg

Lane 5:

NIH/3T3 Membrane extract at 30 µg

Lane 6:

HepG2 Membrane extract at 30 µg

Lane 7:

HeLa Membrane extract at 30 µg

Lane 9:

T-47D Membrane extract at 30 µg

Secondary

All lanes:

HRP-conjugated Goat anti-Rabbit IgG (H+L) Secondary Antibody at 1/2500 dilution

Observed band size: 29 kDa

true

• WB

Supplier Data

Western blot - Anti-YWHAE antibody [RP23040165] (AB308051)

Knockdown of 14-3-3 epsilon/YWHAE was achieved by transfecting SH-SY5Y with 14-3-3 epsilon specific siRNAs. Western blot analysis was performed using modified whole cell extracts from the 14-3-3 epsilon knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with ab308051 at 0.25ug/ml and Goat anti-Rabbit IgG (H+L) Recombinant Secondary Antibody, HRP conjugate at 1 : 4000 dilution.

All lanes:

Western blot - Anti-YWHAE antibody [RP23040165] (ab308051) at 0.25 µg/mL

Lane 1:

Untransfected SH-SY5Y cells

Lane 2:

non-targeting scrambled siRNA transfected SH-SY5Y cells

Lane 3:

14-3-3 epsilon knockdown SH-SY5Y cells

Secondary

All lanes:

HRP-conjugated Goat anti-Rabbit IgG (H+L) Recombinant Secondary Antibody at 1/4000 dilution

Observed band size: 30 kDa

false