Affinity purification–mass spectrometry of TaRIPK-associated fungal proteins
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ABSTRACT: OE-TaRIPK-#2 plants were inoculated with FGJQ. To enhance fungal infection efficiency, an additional set of OE-TaRIPK-#2 plants was treated with BSMV:TaRBOHD prior to FHGQ inoculation. At 9 dpi, total proteins were extracted from each group using radio immunoprecipitation assay (RIPA) buffer and desalted using Zeba™ spin desalting columns. The purified proteins were incubated with 30 μL myc-conjugated agarose beads in binding buffer at 4°C overnight under gentle rotation. After centrifugation at 2,000 x g for 3 min at 4°C, the pellet was washed sequentially with Buffer I (2% sodium dodecyl sulfate (SDS), Buffer II (50 mM HEPES, 500 mM NaCl, 1 mM EDTA, 0.1% deoxycholic acid, and 1% Triton X-100), and Buffer III (10 mM Tris–HCl, 250 mM LiCl, 1 mM EDTA, 0.1% deoxycholic acid, and 1% IGEPAL® CA-630. Following six washes with 50 mM ammonium bicarbonate, proteins were digested with trypsin (1:50) at 37°C overnight.
ORGANISM(S): Triticum Aestivum
SUBMITTER:
Zaifeng Li
PROVIDER: PXD075184 | iProX | Wed Mar 04 00:00:00 GMT 2026
REPOSITORIES: iProX
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