Plant/Algal cell antibodies / Membrane transport system / Vacuolar membrane

product information

Na(+)/H(+) exchanger 1 protein is involved in exchange of protons for cations such as sodium and potassium across membraes. Localized in tonoplast, possibily also to ER and Golgi. Alternative name: NHE-1, Na(+)/H(+) exchanger 1

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana NHX protein  Q68KI4

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

0.1 % sodium azide is added as preservative. For antibody re-suspending information check the tube lable.

Protocol for vacuolar membrane isolation can be found here.

application information

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

59.5 | 45 kDa

Arabidopsis thaliana

dictos including: Gossypium hirsutum, Ricinus communis, Vitis vinifera, monocots including: Oryza sativa, Zea mays, trees: Populus euphratica

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.

Manufacture by Operon Biotechnologies.

to be added when available

application example

1 µg and 10 µg of crude membrane fraction/lane from Arabidopsis thaliana  (left panel) and 1,5 and 10 µg of crude membrane fraction/lane Raphanus sativus L. (right panel) 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-NHX antibodies (AS09 484, 1:1000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.