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Both antibodies were found to label rod outer segments only, leaving rod inner segments unlabeled (see vertical sections in Figure 4A,4B).
Anti-rhodopsin antibody (dilution 1∶250) and PNA lectin from Arachis hypogae (FITC-conjugated peanut agglutinin, dilution 1∶50) were incubated overnight at 4°C to label rod and cone photoreceptor cells respectively.
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We double-labeled retinal sections with an anti-Chx10 antibody, a selective marker for differentiated rod and cone BCs, and an anti-protein kinase Cα (anti-PKCα) antibody that selectively labels rod BCs (Green et al. 2003).
While both antibodies clearly labeled rods in both dunnart and wallaby, the particular rod opsin epitopes might not be detectable or accessible.
The SNAP tag labeled rods behaved like their S tag labelled counterparts during expression and purification from E.coli extracts.
This labeling of rod terminals is unlikely to be a fixation artifact, since the sections were lightly fixed and labeling was specific to synaptic terminals.
The GFP expressing cells can be purified by FACS technology to contain nearly 90% photoreceptors, as assessed with subsequent labeling with rod and cone markers.
As CrxGFP labels both rod and cones cells, use of this transgene allowed us to investigate the integration efficiency of both photoreceptor types simultaneously, using donor cells isolated from different ontogenetic stages.
We provide methods for purifying SNAP tag labeled septin rods in high yield and purity.
Filament formation and binding assays confirmed that the labeled septin rods display native-like properties.
Kim Richman, a lawyer who spoke at a seminar during the expo, called "all natural" labels "a lightning rod" that can lead to costly and time-consuming lawsuits no matter what's in the package.
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