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

tested applications

ELISA (ELISA), western blot (WB)

related products

collection of antibodies to plasma membrane proteins

AS09 488 | anti-PIP2;1 | aquaporin PIP2;1

AS09 489 | anti-PIP1;1, PIP1;2, PIP1;3 | aquaporins

AS09 491 | anti-PIP2;1,PIP2;2,PIP2;3 |plasma membrane intrinistic protein 2-1,2-2, 2-3

additional information

0.1 % sodium azide is added as preservative. For antibody re-suspending information check the tube lable.

Antibodies will detect target protein in a few µg of a crude preparation loaded per well. If purified preparations of vacuolar and plasma membranes are used, one µg load per well should be sufficient.

Protocol for plant plasma membrane isolation can be found here.

application information
recommended dilution		1: 8000 (ELISA), 1: 2000 with standard ECL (WB)
expected \| apparent MW		30 \| 28 kDa (Raphanus sativus)
confirmed reactivity		Arabidopsis thaliana, Raphanus sativus
predicted reactivity		dicots including: Brassica sp.
not reactive in		no confirmed exceptions from predicted reactivity known in the moment
additional information		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.
selected references		to be added when available

application example

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.

western blot using anti-PIP2;2 antibodies

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