PIP1;1, PIP1;2, PIP1;3, PIP1;4, PIP1;5 | Aquaporins

KLH-conjugated synthetic peptide derived from N terminus of Raphanus sativus PAQ1O80368. Chosen peptide is conserved in PIP1;1, PIP1;2, PIP1;3 N-terminus of Raphanus sativus and in  all 5 isoforms of Arabidopsis thaliana coded by:AT3G61430.1 (PIP1;1), AT2G45960.3 (PIP1;2), AT1G01620.1 (PIP1;3), AT4G00430.1 (PIP1;4), AT4G23400.1 (PIP1;5)

30.66 | 28 kDa (Raphanus sativus)

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-PIP1s ntibodies (diluted in 1: 500 (left panel) and 1: 1000 (right panel), AS09 489, and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.

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. PIP1;1-PIP1;5 isoforms can be detected in most tissues.

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.

Antibody reacts also with PIP2 isoforms in Zea mays.

collection of antibodies to plasma membrane proteins

AS09 488 | Anti-PIP2;1 | aquaporin PIP2;1, rabbit antibodies

AS09 490 | Anti-PIP2;2 | plasma membrane aquaporin 2b, rabbit antibodies

AS09 491 | Anti-PIP2;1,PIP2;2,PIP2;3 |plasma membrane intrinistic protein 2-1,2-2, 2-3, rabbit antibodies

Plant protein extraction buffer

Secondary antibodies

PIPs proteins are aquaporins which facilitate the transport of water and small neutral molecules across cell membrane. PIP1;2 is also designated as PIP1B.