PIP2;1+PIP2;2 | Aquaporin PIP2;1+PIP2;2

AS09 488 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, O. sativa, R. sativus

Product Information

Immunogen

KLH-conjugated synthetic peptide derived from Raphanus sativus PAQ2 O80369

Host Rabbit

Clonality Polyclonal

Purity Serum

Format Lyophilized

Quantity 100 µl

Reconstitution For reconstitution add 100 µl of sterile water.

Storage Store lyophilized/reconstituted at -20°C; once reconstituted 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 lyophilized material adhering to the cap or sides of the tubes.

Tested applications ELISA (ELISA), Western blot (WB)

Recommended dilution 1 : 8000 (ELISA), 1 : 1000 (WB)

Expected | apparent MW

30.34 | 28 kDa (Raphanus sativus)

Reactivity

Confirmed reactivity Arabidopsis thaliana, Oryza sativa, Raphanus sativus

Predicted reactivity Brassica napus, Gossypium mexicanum

Not reactive in Hordeum vulgare

Application examples

Application example

western blot using anti-PIP2;1 antibodies

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-PIP2;1 antibodies (AS09 488, 1:1000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.

Additional information

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.

Triton X-100 should not be included in the protein extraction buffer, when cell organelles or membrane proteins must be separated from soluble proteins. Because, Triton X breaks membrane structure and solubilizes most membranes proteins. Furthermore, it should be noted that Triton X at high concentrations binds SDS and mask the detergent effect of SDS for SDS-PAGE. Also, micelles of Triton X behave as a large complex with molecular mass of 90 kDa at high concentrations in SDS-PAGE.

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.

Peptide used to elicit this antibody is also conserved in Arabidopsis thaliana PIP2;1,PIP2;2, PIP2;3 and this antibody reacted also with PIP2;2 in experiments with knock out plants.

Related products

collection of antibodies to plasma membrane proteins

AS09 489 | anti-PIP1;1, PIP1;2, PIP1;3 | aquaporins

AS09 490 | anti-PIP2;2 | plasma membrane aquaporin 2b

AS09 491 | anti-PIP2;1,PIP2;2,PIP2;3 |plasma membrane intrinistic protein 2-1,2-2, 2-3

Plant protein extraction buffer

Secondary antibodies

Background

PIPs proteins are aquaporins which facilitate the transport of water and small neutral molecules across cell membrane.

Product citations

Selected references Markus G. et al., (2004). Arabidopsis Immunophilin-like TWD1 Functionally Interacts with Vacuolar ABC Transporters.Mol. Biol. Cell. 15, 3393-3405.