Plant/Algal cell biology / Conifers

product information

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

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana  P25818

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 493 | 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.

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.

Protocol for isolation of plant vacuolar membranes can be found here.

application information

1: 8000 (ELISA), 1: 2000 with standard ECL (WB)

25.6 | 24 kDa (Arabidopsis thaliana)

Arabidopsis thaliana

dictos including: Brassica oleracea, Gossypium mexicanum, Pisum sativum, Ricinus communis, Vitis vinifera, monocots including: Oryza sativa, trees: Picea abies, Popoulus 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.

Manufactured by Operon Biotechnologies.

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. 25:5814-5820.

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.