Rabbit anti p src tyr416

Cell lysis, SDS-polyacrylamide gel electrophoresis, protein transfer and western blot were performed similarly as before^{31 (link)}. Dual Color Protein Standard (Bio-Rad, Hercules, CA) was used to determine molecular weight. Antibodies used included rabbit anti-phospho-FAK (Tyr397), (Cell Signaling Technology, Danvers, MA), mouse anti-FAK clone 4.47 (EMD Millipore, Darmstadt, Germany), rabbit anti-FoxM1 (Novus Biologicals, Littleton, CO), rabbit anti-Phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204) (D13.14.4E) (Cell Signaling Technology), rabbit anti-ERK total (Cell Signaling Technology), rabbit anti-phospho-AKT (Ser473) (D9E) (Cell Signaling Technology), mouse anti-GAPDH (EMD Millipore), rabbit anti-AKT total (Cell signaling Technology), rabbit anti p-Src (Tyr416) (Cell Signaling Technology) and mouse anti-total Src (Cell signaling Technology).

Images were collected using a Leica DMI 4000B fluorescence microscope. To detect caveolin-1, CCR1, CCR2, and CCR3, cells isolated as described above were cultured overnight in 6-well tissue culture plates (1 × 10⁶ cells per well) on coverslips in RPMI 1640/20 % FCS. Cells were then fixed and permeabilized, labeled with appropriate primary and secondary antibodies, and counterstained with the nuclear stain DAPI.
Immunohistochemistry of human lung tissue sections was performed as described [11 (link)]. Briefly, paraffin sections were stained with primary antibodies, appropriate AlexaFluor647- or AlexaFluor555-conjugated secondary antibodies and the nuclear stain DAPI (Invitrogen, Carlsbad, CA). Images were collected using a Leica DMI 4000B fluorescence microscope. Primary antibodies were: rabbit anti-CCR1 (Thermo Fisher Scientific, Rockford, IL, USA; PA1-21629), rabbit anti-CCR2 (Abcam, Cambridge, MA, USA; ab32144), rabbit anti-CCR3 (Abcam, Cambridge, MA, USA; ab36827), rabbit anti-MCP-1 (Abcam, Cambridge, MA, USA; ab 9669), rabbit anti-MCP-3 (Santa Cruz Biotechnology, Santa Cruz, CA, USA; SC-374002), rabbit anti-pSrc-Tyr416 (Cell Signaling Technology, Inc., Danvers, MA, USA; #2101S), and rabbit anti-pLyn-Tyr507 (Cell Signaling Technology, Inc., Danvers, MA, USA; 04–375).