SIP1 | Small basic intrinsic protein 1-1

KLH-conjugated synthetic peptide conserved in Arabidopsis thaliana SIP1;1 Q9M8W5, At3g04090

25.7 | 25 kDa

Application example

Crude membrane fraction/lane from Arabidopsis thaliana stems (1,4) and crude membrane fraction from yeast cells expressing SIP1;1 (2,5) and SIP1;2 (3,6) 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-AtSIP1;1 antibodies (AS09 495, 1:1000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.

Peptide used to elicit anti-SIP1;1 antibodies have been incubated with antibodies - right panel.

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.

Anti-SIP1;1 antibodies will not recognize SIP2;1 protein.

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.

AS09 496 | anti-AtSIP1;1

collection of antibodies to endomembrane system

Plant protein extraction buffer

SIP protein functions as a water channel to facilitate the transport of water across cell membrane. It is mainly localized to ER. Alternative name: small basic intrinistic protein 1-1