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CBMs as Probes to Explore Plant Cell Wall Heterogeneity Using Immunocytochemistry.

Methods Mol Biol.. 2017-04; 
Badruna L,Burlat V,Montanier CY.
Products/Services Used Details Operation
... 3. CBM3a (homemade production with an Histidine-Tag (see Note 2 )). 4. Mouse monoclonal anti-Histidine Tag antibody (A00186-100, GenScript). 5. Goat anti-mouseIgG-FITC antibodies (Sigma, F2012). 2.4 Sample Preparation Materials. 1. Dry oven. 2. Vacuum chamber. 3. ...

摘要

Immunocytochemistry is a widely used technique to localize antigen within intact tissues. Plant cell walls are complex matrixes of highly decorated polysaccharides and the large number of CBM families displaying specific substrate recognition reflects this complexity. The accessibility of large proteins, such as antibodies, to their cell wall epitopes may be sometimes difficult due to steric hindrance problems. Due to their smaller size, CBMs are interesting alternative probes. The aim of this chapter is to describe the use of CBM as probes to explore complex polysaccharide topochemistry in muro and to quantify enzymatic deconstruction.

关键词

Enzymatic hydrolysis; Fluorescence; Immunocytochemistry; Microscopy; Wheat straw section