Plant/Algal cell antibodies / Membrane transport system / Endomembrane system

product information

SIP protein functions as a water channel to facilitate the transport of water across cell membrane. It is mainly localized to ER. Alternative name: small basic intrinistic protein 1-1

KLH-conjugated synthetic peptide conserved in Arabidopsis thaliana SIP1;1 Q9M8W5

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)

AS09 496 | anti-AtSIP1;1

collection of antibodies to endomembrane system

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.

Anti-SIP1;1 antibodies will not recognize SIP2;1 protein.

Method for plant ER isolation is available here.

application information

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

25.7 | 25  kDa

Arabidopsis thaliana

to be determined

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.

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. 579:5814-5820.

application example

crude membrane fraction/lane from Arabidopsis thaliana  stems (1,4) and crude membrane fraction from yeast cells expressing SIP1;1 (2,5) and SIP1;2 (3,6) 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-AtSIP1;1 antibodies (AS09 495, 1:1000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.

Peptide used to elicit anti-SIP1;1 antibodies have been incubated with antibodies - right panel.