TIP1 | Tonoplast intrinsic protein 1-1

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana UniProt: P25818,TAIR:At2g36830

25.6 | 24 kDa (Arabidopsis thaliana)

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-TIP1;1 antibodies (AS09 482, 1:1000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.

TIP1;1 usually shows a faint band of dimeric form at 42 kDa in addition to the major monomoer band of 23 kDa even in the presence of SDS.

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.

TIP1;1 usually shows a faint band of dimeric form at 42 kDa in addition to the major monomoer band of 23 kDa even in the presence of SDS.

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.

TIP1;1 is a water channel protein needed for the transport of water, diffusion of amino acids and peptides. Alternative name: Tonoplast intrinsic protein 1-1, gamma-tonoplast intrinsic protein, Gamma-TIP, aquaporin TIP, Tonoplast intrinsic protein, root-specific RB7