PIP (PIP1;1, PIP1;2, PIP1;3, PIP1;4, PIP1;5) | Aquaporins

AS09 487 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, J. curcas, P. nigra, P. trichocarpa, R. sativus

Product Information

Immunogen

KLH-conjugated synthetic peptide conserved in Arabidopsis thaliana: PIP1;1 UniProt: P61837,At3g61430 PIP1;2 UniProt: Q06611,TAIR: At2g45960 PIP1;3 UniProt: Q08733,TAIR: At1g01620 , PIP1;4 UniProt: Q39196,TAIR: At4g00430, PIP1;5 UniProt: Q8LAA6 TAIR:At4g23400

Host Rabbit

Clonality Polyclonal

Purity Affinity purified serum in PBS, pH 7.4

Format Lyophilized

Quantity 50 µg

Reconstitution For reconstitution add 50 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 Western blot (WB)

Recommended dilution 1 : 1000 (WB)

Expected | apparent MW

30.68 | 28 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana, Brassica sp., Jatropha curcas L. cv. Biji Jarak , Mesembryantheum crystallinum, Populus nigra, Populus trichocarpa, Raphanus sativus, Thellungiella salsuginea

Predicted reactivity Brassica sp., Hordeum vulgare, Juglans regia, Oryza sativa, Populus tremula, Triticum aestivum, Vicia faba
Species of your interest not listed? Contact us

Not reactive in Fragaria sp., Spinacia oleracea

Application examples

Application examples Application information

10 µg of Arabidopsis thaliana tonoplast fraction (1), Thellungiella salsuginea tonoplast fraction (2), Mesembryanthemum crystallinum tonoplast fraction (3), Nicotiana tabacum tonoplast fraction (4), Arabidopsis thaliana plasma membrane fraction (5), Thellungiella salsuginea plasma membrane fraction (6), Mesembryanthemum crystallinum plasma membrane fraction (7), Arabidopsis halleri microsome fraction (8), Brassica sp. microsomal fraction (9) were extracted with Protein Extraction Buffer PEB (AS08 300). Samples were diluted with 1X sample buffer (NuPAGE LDS sample buffer (Invitrogen) supplemented with 50 mM DTT and heat at 70°C for 5 min and keept on ice before loading. Protein samples were separated on 4-12% Bolt Plus gels, LDS-PAGE and blotted for 70 minutes to PVDF using tank transfer. Blots were blocked immediately following transfer in 2% blocking reagent or 5% non-fat milk dissolved in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 10 000 (in blocking reagent) for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, and then washed 1x15 min and 3x5 min with TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, recommended secondary antibody AS09 602,Agrisera) diluted to 1:25 000 in blocking reagent for 1h at room temperature with agitation. The blots were washed as above. The blot was developed for 5 min with chemiluminescence reagent according to manufacture instructions. Images of the blots were obtained using a CCD imager (VersaDoc MP 4000) and Quantity One software (Bio-Rad). Exposure time was 30 seconds.

The background at the top of the membrane can be optimized.

Courtesy of Dr. Rosario Vera, UNAM, Mexico

Additional information

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.

Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel.

Related products

collection of antibodies to plasma membrane proteins

AS09 487-ALP | Anti-PIP (PIP1;1, PIP1;2, PIP1;3, PIP1;4, PIP1;5) aquaporins, ALP-conjugated (40 µg), rabbit antibodies
AS09 487-HRP | Anti-PIP (PIP1;1, PIP1;2, PIP1;3, PIP1;4, PIP1;5) aquaporins, HRP-conjugated (40 µg), rabbit antibodies

Plant protein extraction buffer

Secondary antibodies

Background

PIPs proteins are aquaporins which facilitate the transport of water and small neutral molecules across cell membrane. Alternative names: PIP1-1, PIP1-2, PIP1-3, plasma membrane intrinistic protein 1a, 1b, 1c, PIP1a, PIP1b, PIP1c, plasma membrane aquaporin-1, transmembrane protein A, TMP-A, AthH2, transmembrane protein B, TMP-B,