grasp mouse anti gfp  (NeuroMab)

Journal: eNeuro

Article Title: Wakefulness Is Promoted during Day Time by PDFR Signalling to Dopaminergic Neurons in Drosophila melanogaster

doi: 10.1523/ENEURO.0129-18.2018

Figure Lengend Snippet: Details of fly stocks used in the study.

Article Snippet: Fixed brains were blocked in 10% horse serum for 1 h at room temperature and 6–9 h at 4°C, followed by incubation with cocktail containing primary antibodies for 48 h. The primary antibodies used were anti-GFP (chicken, 1:2000, for GFP labeling and CaLexA measurements, Invitrogen #A10262, RRID: AB_2534023 ), anti-PDF (mouse, 1:5000, DSHB, PDF C7, RRID: AB_760350 ), anti-MYC (mouse, 1:1000, Cell Signaling Technology, #9B11, RRID: AB_2148465 ), anti-TH (rabbit, 1:1000, Invitrogen #P21962, RRID: AB_2539844 ), anti-GFP (mouse, 1:500, for GRASP, Sigma-Aldrich #G6539, RRID: AB_259941 ), and anti-PDF (rabbit, 1:30,000, M. Nitabach and T. C. Holmes).

Techniques:

Journal: eNeuro

Article Title: Wakefulness Is Promoted during Day Time by PDFR Signalling to Dopaminergic Neurons in Drosophila melanogaster

doi: 10.1523/ENEURO.0129-18.2018

Figure Lengend Snippet: Anatomic connections between TH + and PDF + neurons. A , Reconstituted GFP (GRASP) signal was detected in brains of flies expressing LexAop CD4::spGFP11 under Pdf LexA control and UAS CD4::spGFP1-10 under TH GAL4 control ( n = 22 hemispheres). GRASP signal colocalized with ascending portion of s-LN v dorsal projections labeled with antibody against PDF. Highlighted region is magnified in the right-most panel. Control flies lacking expression of ( B ) spGFP1-10 ( n = 16 hemispheres) and ( C ) spGFP11 ( n = 16 hemispheres) do not show GRASP signal. Results representative from two independent experiments. Arrowheads indicate non-specific staining. Expression of GFP using TH GAL4 and colabeling PDF ( D , left) and using antibodies against TH and PDF ( D , right) in wild-type flies reveal dopaminergic projection in the vicinity of ascending portion of s-LN v dorsal projection as indicated by asterisks. Scale bar: 20 µm. See also Extended Data .

Article Snippet: Fixed brains were blocked in 10% horse serum for 1 h at room temperature and 6–9 h at 4°C, followed by incubation with cocktail containing primary antibodies for 48 h. The primary antibodies used were anti-GFP (chicken, 1:2000, for GFP labeling and CaLexA measurements, Invitrogen #A10262, RRID: AB_2534023 ), anti-PDF (mouse, 1:5000, DSHB, PDF C7, RRID: AB_760350 ), anti-MYC (mouse, 1:1000, Cell Signaling Technology, #9B11, RRID: AB_2148465 ), anti-TH (rabbit, 1:1000, Invitrogen #P21962, RRID: AB_2539844 ), anti-GFP (mouse, 1:500, for GRASP, Sigma-Aldrich #G6539, RRID: AB_259941 ), and anti-PDF (rabbit, 1:30,000, M. Nitabach and T. C. Holmes).

Techniques: Expressing, Labeling, Staining