Cells were seeded on fibronectin-coated (10 μg/ml, Sigma) glass chamber slides (NUNC) in the presence of 10% FBS for 2.5 h. Cultures were treated with 500 nM of Lck inhibitor (A770041, Axon Medchem) or DMSO vehicle for 2 h and then fixed in 4% formaldehyde. The samples were stained overnight at 4 °C with Phospho-Paxillin antibody (CellSignaling). The following day, secondary antibodies were applied followed by application of Rhodamine RedX Phalloidin (Life Technologies) and Hoechst stain (Life Technologies). Slides were imaged using a Zeiss LSM 750 confocal microscope.
Phospho paxillin antibody
Manufactured by Cell Signaling Technology
The Phospho-Paxillin antibody is a research-use only product that specifically recognizes paxillin when phosphorylated. Paxillin is an adaptor protein involved in cell adhesion and signaling pathways.
Automatically generated - may contain errors
Lab products found in correlation
Hoechst stain, by Thermo Fisher Scientific (1 mentions)
Glass chamber slide, by Thermo Fisher Scientific (1 mentions)
Lsm 750 confocal microscope, by Zeiss (1 mentions)
Fibronectin coated, by Merck Group (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Thermo Fisher Scientific (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Goat serum, by Merck Group (1 mentions)
2 protocols using phospho paxillin antibody
1
Paxillin Phosphorylation Visualization
2
Immunofluorescence Staining of Phospho-Paxillin
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Cells were fixed for 15 min in 4% PFA (Sigma), then blocked and permeabilized for 10 min in 0.05% goat serum (Sigma) and 0.3% Triton X-100 (Sigma), followed by incubation with 1 : 50 phospho-paxillin antibody (2541 Cell Signaling Technology) in 0.3% Triton X-100 and 0.002% BSA (Gibco) overnight at 4°C. Samples were then incubated with an Alexa Fluor-conjugated secondary antibody or 1 : 800 phalloidin (Abcam) for 90 min, followed by incubation with 2 µg ml−1 33342 Hoechst for 10 min.
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