Anti-YAP1 (phospho S127) antibody [RP23040163]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-YAP1 (phospho S127) Antibody [RP23040163] (AB313464)

For immunofluorescence analysis, PC-3 cells were fixed and permeabilized for detection of endogenous YAP pS (127) using ab313464 (2 ug/ml) and labeled with Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor® 488 conjugate ( 1 : 2000). Panel a) shows representative cells that were stained for detection and localization of YAP pS (127) protein (green), Panel b) is stained for nuclei (blue) using DAPI . Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin ( 1 : 300). Panel d) is a composite image of Panels a, b and c clearly demonstrating cytoplasmic localization of YAP pS (127). Panel e) shows loss of signal by competition with the YAP pS (127) peptide, demonstrating antibody specificity, and panel f) demonstrates no competition with the non-phospho peptide. Panel g) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

• WB

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Western blot - Anti-YAP1 (phospho S127) Antibody [RP23040163] (AB313464)

A 54 kDa band corresponding to YAP (pS127) was observed across the cell lines tested. Known quantity of protein samples were electrophoresed 4-12% Bis-Tris gel. 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.

All lanes:

Western blot - Anti-YAP1 (phospho S127) Antibody [RP23040163] (ab313464) at 2 µg/mL

Lane 1:

Membrane extracts of U-87 MG at 30 µg

Lane 2:

Membrane extracts of HeLa at 30 µg

Lane 3:

Membrane extracts of NTERA-2 at 30 µg

Lane 4:

Membrane extracts of PC-3 at 30 µg

Secondary

All lanes:

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

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