Comprehensive Cell Wall Polysaccharide Profiling in P. patens

The probes used for labeling cell wall polysaccharides in P. patens were chosen based on an earlier Comprehensive Microarray Polymer Profiling (CoMPP) analysis (Moller et al., 2007 (link)) with some additions (Table 1). Antibodies included anti-homogalacturonan (HG) JIM5, JIM7, LM18, LM19, LM20 (Verhertbruggen et al., 2009 (link)), anti-1-4-β-D-galactan LM5 (Jones et al., 1997 (link)), anti-1-5-α-L-arabinan LM6 (Willats et al., 1998 (link)), anti-1-3-β-D-glucan BS400-4 (Meikle et al., 1991 (link)), anti-xylan LM10 (McCartney et al., 2005 (link)), anti-xyloglucan LM15 (Marcus et al., 2008 (link)), anti-mannan BS400-4 (Pettolino et al., 2001 (link)), and anti-AGP LM2 (Smallwood et al., 1996 (link)) and JIM13 (Knox et al., 1991 (link)). CBMs used for labeling included CBM3a and CBM28 (Blake et al., 2006 (link)). Anti-extensin probes were not tested based on lack of cross-reactivity shown by CoMPP (Moller et al., 2007 (link)). Antibodies designated JIM and LM, along with CBM3A, were obtained from Plant Probes (Leeds, UK) and antibodies designated BS were obtained from Australian Biosupplies (Bundoora, VIC, Australia). CBM28 was a gift of Paul Knox (University of Leeds). Other antibodies used included Alexafluor 488-conjugated anti-mouse and anti-rat (Life Technologies, Grand Island, NY, USA) and mouse anti-His (Sigma–Aldrich, St. Louis, MO, USA).