Molecular recognition reagents are key tools for understanding biological processes and are used universally by scientists to study protein expression, localisation and interactions. Antibodies remain the most widely used of such reagents and many show excellent performance, although some are poorly characterised or have stability or batch variability issues, supporting the use of alternative binding proteins as complementary reagents for many applications. Here we report on the use of Affimer proteins as research reagents. We selected 12 diverse molecular targets for Affimer selection to exemplify their use in common molecular and cellular applications including the (a) selection against various target molecules; (b) modulation of protein function in vitro and in vivo; (c) labelling of tumour antigens in mouse models; and (d) use in affinity fluorescence and super-resolution microscopy. This work shows that Affimer proteins, as is the case for other alternative binding scaffolds, represent complementary affinity reagents to antibodies for various molecular and cell biology applications.DOI:
http://dx.doi.org/10.7554/eLife.24903.001
SUMMARY
Questionnaires completed by 140 growers from 12 allotment sites around Leeds, West Yorkshire, listed a wide range of crop pests. Of the seven crops grown by over 50 per cent of the growers, brassicas, potatoes and carrots were damaged more frequently than expected from the average frequency for all 20 crops, peas and celery less often, and beans and onions at about the expected frequency. The main pests reported for brassicas and potatoes were slugs, and the most common pest of carrots was carrot fly. Further examination of 92 plots revealed a large number of pests; slugs, carrot fly (Psila rosae (F.)), aphids and cabbage root fly (Delia brassicae (Wiedemann)) were the most common. Results for soil samples showed pale potato cyst nematode (Globodera pallida (Stone) Behrens) to be at potentially damaging population levels in 21 out of 33 plots sampled at 10 allotment sites established for more than five years and in one of six plots sampled at two newly formed allotment sites. Soil sampling for carrot fly at eight allotment sites revealed widespread overwintering populations of about 1·5 larvae and pupae/100 ml of soil. Baiting for slugs on one plot indicated that a large population of these animals was present.
Pesticides were not used by 22 of the 140 growers in the survey. The most widely favoured chemicals with at least 20 users each were a disinfectant, slug pellets, organophosphates and calomel.
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