PIP2;3 | Aquaporin, plasma membrane intrinistic protein 2-3

AS09 506 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Oryza sativa, Zea mays

Product Information

Immunogen

KLH-conjugated peptide derived from Oryza sativa PIP2;3 Q7XUA6

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), Immunogold (IG), Western blot (WB)

Recommended dilution 1 : 8000 (ELISA), 1 : 100 (IG), 1 : 2000 (WB)

Expected | apparent MW

31.9 | 26-29 kDa

Reactivity

Confirmed reactivity Oryza sativa, Zea mays

Not reactive in

Arabidopsis thaliana, Hordeum vulgare

Application examples

Application example

western blot detection using anti-PIP2;1

0.5 µg of crude membrane fraction/lane from Oryza sativa L. cv. Akitakomachi leaf blade and root 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 506, 1:1000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.

Peptide used to elicti this antibody has been added to primary antibody incubation (right panel, + +) and therefore specific signal has been depleted in this neutralization/immunocompetition assay.

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.

For immunogold technique, secondary antibodies conjugated with coloidal gold, 10 nm particle size were used.

Antibody works very well on samples from roots. 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.

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.

Related products

collection of antibodies to plasma membrane proteins

AS09 505 | anti-PIP1s | aquaporin

AS09 507 | anti-PIP2;1 | aquaporin, plasma membrane intrinistic protein 2-1

AS09 508 | anti-PIP2;5 | aquaporin, plasma membrane intrinistic protein 2-5

Plant protein extraction buffer

Background

PIP2;3 is an aquaporin which facilitate the transport of water and small neutral molecules across cell membrane. Alternative names: plasma membrane intrinsic protein 2-3, OsPIP2;3

Product citations

Selected references Sakurai et al. (2008). Tissue and cell-specific localization of rice aquaporins and their water transport activities- Plant Cell Physiol. 49: 30-39.