Anti-PIP2;2 | Plasma membrane aquaporin 2b

Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel.Antiboy will weakly react with PIP2;1 and theoretically with PIP2;3, however this protein is very scarcely expressed in plant tissues. The mRNA content is 1 % of that for PIP2;2, therefore reactivity to PIP2;3 was neglegible in our experiments.Triton X-100 should not be included in the protein extraction buffer, when cell organelles or membrane proteins must be separated from soluble proteins. Because, Triton X breaks membrane structure and solubilizes most membranes proteins. Furthermore, it should be noted that Triton X at high concentrations binds SDS and mask the detergent effect of SDS for SDS-PAGE. Also, micelles of Triton X behave as a large complex with molecular mass of 90 kDa at high concentrations in SDS-PAGE.

Brilladaet al. (2020). Exocyst subunit Exo70B2 is linked to immune signalling and autophagy, The Plant Cell, 2020;, koaa022, https://doi.org/10.1093/plcell/koaa022

Store lyophilized/reconstituted at -20°C; make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.