GR | Glutathione reductase

Maltose binding protein (MBP) fusion of Zea mays GR, O64409

54 kDa

Application example

10 µg of total protein from (1) Arabidopsis thaliana leaf extracted with Protein Extration Buffer, PEB (AS08 300), (2) Nicotiana tabaccum leaf extracted with PEB, (3) Zea mays extracted with PEB, (4) Hordeum vulgare leaf extracted with PEB, (5) Physcomitrella patens total cell extracted with PEB, (6) Chlamydomonas reinhardtii total cell extracted with PEB,  (7) Synochocystis elongatus total cell extracted with PEB, extracted with PEB, were separated on  4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to nitrocellulose. Blots were blocked in 2 % low fat dry milk in TBS-T (0.1 % Tween 20) for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 2000 for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:30 000 for 1h at room temperature with agitation. The blots were washed as above and developed for  30 seconds  with chemiluminescent detection reagent according the manufacturers instructions.

The nature of 40 kDa cross reaction in this experiment is not known.

Total IgG concentration is 7 µg/ µl

Glutathione reductase (GR, EC 1.6.4.2) is an important enzyme for  plant protection against environmental stress. It functions in plant defense reactions in the conversion of glutathione disulphide to reduced glutathione (GSH).