Western blot analysis of glutamine synthetase on a rat cerebrum lysate (left). Lane 1: 1:5000, lane 2: 1:10,000, lane 3: 1:20,000 dilution of the anti- glutamine synthetase antibody.
Glutamine synthetase staining on a rat cerebrum section (center). Section prepared during antibody development was formalin fixed and paraffin embedded without citrate buffer pretreatment. Note visible staining of astrocytes in the section. Magnification: 40X.
Immunofluorescent staining of SK-N-SH cells (right). Cells were seeded in a 384 well collagen coated Microplates (Material # 353962) at ~ 8,000 cells per well. After overnight incubation, cells were stained using the methanol fix/perm protocol (see Recommended Assay Procedure; Bioimaging protocol link) and the anti- Glutamine Synthetase antibody. The second step reagent was Alexa Fluor® 488 goat anti mouse Ig (Invitrogen)(pseudo colored green). Cell nuclei were counter stained with Hoechst 33342 (pseudo colored blue). The image was taken on a BD Pathway™ 855 or 435 Bioimager System using a 20x objective and merged using the BD AttoVison ™ software. This antibody also stained SH-SY5Y, C6, U87 and U373 cells using both the Triton™ X-100 and methanol fix/perm protocols (see Recommended Assay Procedure; Bioimaging protocol link).