FAK [p Tyr861] Antibody

Western Blot: FAK [p Tyr861] Antibody [NBP2-68146]

Western Blot: FAK [p Tyr861] Antibody [NBP2-68146] - Extracts of 3T3-L1 cells unstimulated or stimulated with 50 ng/ml LIF for 15 minutes (2-4) were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer overnight at 4C, then incubated with JAK2 (pYpY1007/1008) antibody for two hours at room temperature in a 1% BSA-TBST buffer, following prior incubation with: no peptide (1, 2), the nonphosphopeptide corresponding to the phosphopeptide immunogen (3), or the phosphopeptide immunogen (4). After washing, the membrane was incubated with goat F (ab')2 anti-rabbit IgG HRP-conjugate and signals were detected using the Pierce SuperSignal method. The data show that only the phosphopeptide corresponding to JAK2 (pYpY1007/1008) blocks the antibody signal, demonstrating the specificity of the antibody. The data also show the up-regulation of JAK2 (pYpY1007/1008) with LIF treatment in this cell system.

Immunocytochemistry/Immunofluorescence: FAK [p Tyr861] Antibody [NBP2-68146] - Immunofluorescence analysis of FAK [p Tyr861] Antibody was done on 70% confluent log phase A549 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with FAK [p Tyr861] Antibody Rabbit Polyclonal Antibody at 1:250 dilution in 1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Secondary Antibody conjugated to Alexa Fluor 488 at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with a liquid mountant with DAPI. F-actin (Panel c: red) was stained with Alexa Fluor 555 Rhodamine Phalloidin. Panel d is a merged image showing Cell Junctional localization. Panel e is a no primary antibody control. The images were captured at 60x magnification.