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. 2012 Jun 4;159(4):1355–1366. doi: 10.1104/pp.112.199356

Figure 5.

Figure 5.

Coimmunoprecipitation of XXT5HA and XXT2YFP from Arabidopsis protoplasts. A, XXT2YFP was transiently expressed in protoplasts prepared from XXT5HA-expressing plants. Total protein extracts from protoplasts treated with Triton X-100 were applied to an anti-HA agarose column. The elution fractions were treated with or without β-mercaptoethanol (2-ME) SDS-PAGE under reducing and nonreducing conditions, respectively. Proteins were detected by either polyclonal anti-HA or monoclonal anti-GFP antibody (Ab). B, Negative control for immunoprecipitation of XXT5HA and XXT2YFP. Protein extract from wild-type protoplast transiently expressed XXT2YFP was applied to an anti-HA agarose column. The flow-through, wash (W1 and W3), and elution fractions were separated by SDS-PAGE and detected by monoclonal anti-GFP antibodies.