PIP2;2 | Plasma membrane aquaporin 2b
AS09 490 | clonality: polyclonal | host: rabbit | reactivity: Arabidopsis thaliana, Raphanus sativus
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|Recommended dilution||1 : 8000 (ELISA), 1 : 2000 (WB)|
|Expected | apparent MW||
30 | 28 kDa (Raphanus sativus)
|Confirmed reactivity||Arabidopsis thaliana, Fragaria ananassa, Raphanus sativus|
|Predicted reactivity||Brassica sp.|
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
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.
Manufactured by Operon Biotechnology.
to be added when available
1 µg and 10 µg of crude membrane fraction/lane from Arabidopsis thaliana were separated on 12 % SDS-PAGE and blotted 1h to PVDF membrane (40 min. at 10 V using BioRad semidry transfer). Filters were blocked 1h with 5 % low-fat milk powder in TBS-T (0.05% Triton X.100). Membranes were washed 5 times with TBS-T, each time in a fresh polystyrene box and probed with anti-PIP2;2 antibodies (AS09 490, 1:1000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.
PIP proteins usually show a faint band of dimeric form at 55 kDa addition to the major monomer band of 28 kDa even in the presence of SDS.
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