Abstract
Nicotiana benthamiana (N. benthamiana) is a useful model system to transiently express protein at high level. This protocol describes in detail how to transiently express protein in N. benthamiana and how to carry out protein immunoprecipitation in this expression system. This protocol can be broadly used for investigation on protein-protein interaction, protein purification and other related protein assay.
Materials and Reagents
Equipment
Procedure
Representative data
A representative experiment result was shown in the Figure 1. The figure was adapted from the Xu et al. (2014) as listed in the reference. Figure 1. bHLH84-HA co-immunoprecipitates with SNC1-FLAG. Two samples were used in the Co-IP experiment including sample with coexpression of bHLH84 and SNC1-FLAG and sample with expression of only bHLH84, which serves as the negative control.
Notes
Recipes
Acknowledgments
We are grateful for the financial supports from Natural Sciences and Engineering Research Council (NSERC) (Canada) Discovery grant program. The transient protein expression in N. benthamiana was adapted from Van den Ackerveken et al. (1996). The IP protocol was modified from Moffett et al. (2002).
References
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