Journal: Journal of Experimental Botany

Article Title: Trafficking of storage proteins in developing grain of wheat

doi: 10.1093/jxb/ern346

Figure Lengend Snippet: Immunofluorescence double labelling of the outer (A) and inner (B) endosperm of wheat grains at 20 dpa to show the locations of the tagged LMW subunit and HMW subunits. Alexa 568 (red) was conjugated to the anti-mouse secondary antibody recognizing the 9E10 antibody binding to the c-myc tag of the LMW subunit. Alexa 488 (green) was conjugated to an anti-rabbit secondary antibody recognizing the anti-R2-HMW antibody binding to HMW subunits. Micrographs (C) and (D) are single channel images corresponding to micrographs (A) and (B), respectively. The boxes in micrographs (B) and (D) show examples of protein bodies in the same cell which are labelled by the HMW antibody (green) but not by the anti-c-myc antibody (red). The fluorescence micrographs (A) and (C) correspond to the same region of the grain that is shown stained with toluidine blue in D2. Fluorescence micrographs (B) and (D) correspond to those shown stained with toluidine blue in D1.

Article Snippet: The following monoclonal and polyclonal antibodies were used, singly or in combination, diluted in PBS containing 1% BSA 0.05% Tween20: rabbit polyclonal anti-c-myc (A-14) and mouse monoclonal anti-c-myc (9E10) (Autogen Bioclear), both at 1:200 dilution; mouse monoclonal anti-α-gliadin (glia-α-9, 9-68) ( Mitea et al. , 2008 ) diluted 1:400; mouse monoclonal anti-sulphur rich prolamins IFRN0610 ( Brett et al. , 1999 ), diluted 1:400; rabbit polyclonal anti-R2-HMW ( Denery-Papini et al. , 1996 ), diluted 1:100.

Techniques: Immunofluorescence, Binding Assay, Fluorescence, Staining

Journal: Journal of Experimental Botany

Article Title: Trafficking of storage proteins in developing grain of wheat

doi: 10.1093/jxb/ern346

Figure Lengend Snippet: TEM micrographs showing immunogold labelling of the tagged LMW glutenin subunit in the starchy endosperm of 8dpa wheat grain using the anti-c-myc antibody. Labelling can be seen in protein bodies (A, C, D, E, F) and in Golgi vesicles (A, B, C, D, arrows).

Article Snippet: The following monoclonal and polyclonal antibodies were used, singly or in combination, diluted in PBS containing 1% BSA 0.05% Tween20: rabbit polyclonal anti-c-myc (A-14) and mouse monoclonal anti-c-myc (9E10) (Autogen Bioclear), both at 1:200 dilution; mouse monoclonal anti-α-gliadin (glia-α-9, 9-68) ( Mitea et al. , 2008 ) diluted 1:400; mouse monoclonal anti-sulphur rich prolamins IFRN0610 ( Brett et al. , 1999 ), diluted 1:400; rabbit polyclonal anti-R2-HMW ( Denery-Papini et al. , 1996 ), diluted 1:100.

Techniques:

Journal: Journal of Experimental Botany

Article Title: Trafficking of storage proteins in developing grain of wheat

doi: 10.1093/jxb/ern346

Figure Lengend Snippet: Details of sections of starchy endosperm from 12 dpa wheat grain showing labelled protein bodies and unlabelled inclusion bodies (arrows). The antibody used for labelling was the monoclonal 9E10 anti c-myc.

Article Snippet: The following monoclonal and polyclonal antibodies were used, singly or in combination, diluted in PBS containing 1% BSA 0.05% Tween20: rabbit polyclonal anti-c-myc (A-14) and mouse monoclonal anti-c-myc (9E10) (Autogen Bioclear), both at 1:200 dilution; mouse monoclonal anti-α-gliadin (glia-α-9, 9-68) ( Mitea et al. , 2008 ) diluted 1:400; mouse monoclonal anti-sulphur rich prolamins IFRN0610 ( Brett et al. , 1999 ), diluted 1:400; rabbit polyclonal anti-R2-HMW ( Denery-Papini et al. , 1996 ), diluted 1:100.

Techniques: