PIP1;1, PIP1;2, PIP1;3, PIP1;4, PIP1;5 | Aquaporins
AS09 489 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Nicotiana tabacum, Phoenix dactilifera, Raphanus sativus, Zea mays
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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)
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.
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.
PIPs proteins are aquaporins which facilitate the transport of water and small neutral molecules across cell membrane. PIP1;2 is also designated as PIP1B.
Fernández-San Millán et al. (2018). Physiological Performance of Transplastomic Tobacco Plants Overexpressing Aquaporin AQP1 into Chloroplast Membranes. J Exp. Bot. ery148, https://doi.org/10.1093/jxb/ery148.
Pengelly et al. (2014). Transplastomic integration of a cyanobacterial bicarbonate transporter into tobacco chloroplasts. J Exp. Botany, doi:10.1093/jxb/eru156.
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