Plant/Algal cell antibodies / Membrane transport system / Vacuolar membrane

product information

TIPs belong to MIP/aquaporin protein family.Alternative names: Aquaporin TIP1-1,gamma-tonoplast intrinsic protein, gamma-TIP, aquaporin TIP, gamma-tonoplast intrinsic protein, gamma-TIP, aquaporin TIP, tonoplast intrinsic protein, root-specific RB7, gamma-tonoplast intrinsic protein 2, gamma-TIP2, salt stress-induced tonoplast intrinsic protein

KLH-conjugated synthetic peptide conserved in Raphanus sativus TIP1;1 and TIP1;2 (protein accesion number available in Suga et al. 2001). Peptide is also conserved in Arabidopsis thaliana TIP1-1 P25818, TIP1-2 Q41963

store at -20°C; make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tubes.

ELISA (ELISA), western blot (WB)

collection of antibodies to tonoplast proteins

AS09 492 | anti-TIP1;1, TIP1;2 | tonoplast intrinistic protein 1-1, 1-2 (gamma)

AS09 494 | anti-TIP2;1 (delta) | Delta-VM23

AS09 510 | anti-TIP2;1 | tonoplast intrinistic protein 2-1

AS09 511 | anti-TIP2;2 | tonoplast intrinistic protein 2-2 (C-terminal)

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.

Protocol for isolation of vacuolar membranes can be found here.

application information

1: 8000 (ELISA), 1: 1000 with standard ECL (WB)

25.8| 23  kDa

Arabidopsis thaliana, Raphanus sativus

dicots including: Brassica napus, Gossypium hirsutum, Ricinus communis, monocots including: Hordeum vulgare, Oryza sativa, trees: Populus trichocarpa,

no confirmed exceptions from predicted reactivity known in the moment

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.

Suga et al. (2001). Specificity of the accumulation of mRNAs and proteins of the plasma membrane and tonoplast aquaporings in radish organs. Planta 212:294-304. 

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-TIP1s antibodies (AS09 473, 1:1000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.

TIP proteins usually show a faint band of dimeric form at 42 kDa in addition to the major monomer band of 23 kDa even in the presence of SDS.