mouse monoclonal anti green fluorescent protein gfp  (Novus Biologicals)

Journal: Investigative Ophthalmology & Visual Science

Article Title: Differential Eye Expression of Xenopus Acyltransferase Gnpat and Its Biochemical Characterization Shed Light on Lipid-Associated Ocular Pathologies

doi: 10.1167/iovs.64.5.17

Figure Lengend Snippet: The amino end of Gnpat is necessary for a functional enzyme. ( A ) A double knock-out sct1 Δ gpt2 Δ strain expressing GPT2 under the GAL promoter in a LEU2 plasmid was transformed with a URA3- based plasmid containing wild-type gnpat or truncated Δ129N- gnpat . Four independent transformants were grown in selective –ura/–leu plates containing galactose and then re-streaked on galactose or glucose plates and grown for 3 days at 30°C. ( B ) Expression of Gnpat-GFP, ΔN-Gnpat-GFP, and GFP in wild-type cells analyzed by western blotting using anti-GFP antibodies in lysates and soluble and membrane preparations obtained from subcellular fractionation. Membranes stained with Ponceau Red dye after transfer are shown for the loading control.

Article Snippet: For regular immunohistochemistry, 12-μm sections from stage 39 electroporated tadpoles were stained with an Invitrogen mouse anti-green fluorescent protein (GFP) monoclonal (3E6) antibody (1/1000, A11122; Thermo Fisher Scientific) and a BD Pharmingen rabbit polyclonal anti-activated capase-3 antibody (1/500, 559565; BD Biosciences, Franklin Lakes, NJ, USA), followed by the corresponding Alexa Fluor-tagged secondary antibody.

Techniques: Functional Assay, Knock-Out, Expressing, Plasmid Preparation, Transformation Assay, Western Blot, Membrane, Fractionation, Staining, Control