Images for product: anti-MAP kinase p38 (p38) (pThr180) antibody
Western blot analysis for p38 MAPK (pT180/pY182). HeLa cells (Human cervical epitheloid carcinoma, ATCC CCL-2) were either left untreated (lane 1) or treated (lane 2) with 25 µg/ml anisomycin, an antibiotic and protein synthesis inhibitor known to activate signal transduction pathways, for 15 minutes at 37°C. The top panel was probed with a mouse anti-p38alpha antibody and the bottom was probed with the mouse anti-p38 MAPK (pT180/pY182) antibody at a 1:2500 dilution. The target band in each panel is observable at 38-42 kD.
Immunofluorescent staining of HeLa cells. HeLa cells (Human cervical epitheloid carcinoma, ATCC CCL-2) were seeded in a 96-well imaging plate at ~ 10,000 cells per well. After overnight incubation, cells were either left untreated (left image) or exposed to anisomycin for 30 minutes (right image). After treatment cells were stained using the alcohol perm protocol and the mouse anti-p38 MAPK (pT180/pY182) antibody. The second step reagent was Alexa-Fluor® 488 goat anti-mouse IgG (Invitrogen). Phospho p38 staining is pseudo-colored green, nuclei were stained with Hoechst 33342 and are pseudo-colored blue. Note the lack of staining for phospho-p38 in the untreated cells (left) and the nuclear/cytoplasmic staining in the stimulated cells (right). The images were captured using a 20x objective. This antibody also stained U-2 OS (ATCC HTB-96) cells and can be used with either the Triton™ X-100 or alcohol perm protocols.