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

AS09 490 | anti-PIP2;2 | plasma membrane aquaporin 2b

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 plasma membrane preparation can be found here.

application information
recommended dilution		1: 8000 (ELISA), 1: 1000 with standard ECL (WB)
expected \| apparent MW		30.34 \| 28 kDa (Raphanus sativus)
confirmed reactivity		Arabidopsis thaliana, Oryza sativa, Raphanus sativus
predicted reactivity		dicots including: Brassica napus, Gossypium mexicanum,
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. Diluted antibody solution can be used 2 to 3 times within one month if it contains 0.1 % sodium azide as preservative and is stored at -20ºC to -80ºC. Peptide used to elicit this antibody is also conserved in Arabidopsis thaliana PIP2;1,PIP2;2, PIP2;3 and this antibody reacted also with PIP2;2 in experiments with knock out plants. Manufactured by Operon Biotechnology
selected references		Markus G. et al., (2004). Arabidopsis Immunophilin-like TWD1 Functionally Interacts with Vacuolar ABC Transporters.Mol. Biol. Cell. 15, 3393-3405.

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;1 antibodies (AS09 488, 1:1000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.

western blot using anti-PIP2;1 antibodies

||| For applications or usage on species others than stated above Agrisera offers a payment-after-testing option. To learn more about this or for any questions on this product, please use the LiveChat option in the left menue bar or contact us at support@agrisera.com