SIP2 | Small basic intrinsic protein 2-1
AS09 496 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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crude membrane fraction/lane from Arabidopsis thaliana stems (1,3) and crude membrane fraction from yeast cells expressing SIP2;1 (2,4) 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-AtSIP2;1 antibodies (AS09 496, 1:1000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.
Peptide used to elicit anti-SIP2;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.
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.
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 2-1
Ishikawa et al. (2005). Novel type aquaporin SIPs are mainly localized to the ER membrane and show cell-specific expression in Arabidopsis thaliana. FEBS Lett. 579:5814-5820.
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