Analytical implications of different methods for preparing plant cell wall material.

• Preparation of an Alcohol Insoluble Residue (AIR) is an important first step in the analysis of cell wall carbohydrates. • Variation in the protocols used for AIR preparation impacts the downstream extractability and detection of cell wall components. • Carbohydrate microarrays were used to analyse carbohydrate extractability. • 70-CHCl3/MeOH L protocol can be considered a reasonable starting point for optimisation of carbohydrate extraction. Almost all plant cells are surrounded by a wall constructed of co-extensive networks of polysaccharides and proteoglycans. The capability to analyse cell wall components is essential for both understanding their complex biology and to fully exploit their numerous practical applications. Several biochemical and immunological techniques are used to analyse cell walls and in almost all cases the first step is the preparation of an alcohol insoluble residue (AIR). There is significant variation in the protocols used for AIR preparation, which can have a notable impact on the downstream extractability and detection of cell wall components. To explore these effects, we have formally compared ten AIR preparation methods and analysed polysaccharides subsequently extracted using high-performance anion exchange chromatography (HPAEC-PAD) and Micro Array Polymer Profiling (MAPP). Our results reveal the impact that AIR preparation has on downstream detection of cell wall components and the need for optimisation and consistency when preparing AIR. [ABSTRACT FROM AUTHOR]