Anti-GLO1 | (S)-2-hydroxy-acid oxidase GLO1

1 - 50 ug of Arabidopsis thaliana whole leaf extract
2 - 50 ug of Arabidopsis thaliana mutant

15 µg/well of total protein extracted from whole leaves of Aranidopsis thaliana, in 2X laemmli buffer and denatured with exact buffer components at 70°C/10 min. Samples were separated on tris/glycine 11 % acryl gel and blotted for 2h at 170mA on PVDF (Millipore), using: wet transfer. Blot was blocked with 5 % milk for 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1000 ON/4°C with agitation in TBST+5% milk. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 3 times for 10 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1: 10 000 in for 1h/RT with agitation (tested provided secondary). The blot was washed as above and developed with a following chemiluminescent detection reagent: Agrisera Bright (mid picogram detection range).
Exposure time was 30sec.

Courtesy of Dr. Marion Clavel, Gregor Mendel Institute of Molecular Plant Biology, Austria

(S)-2-hydroxy-acid oxidase GLO1 is involved in a subpathway of photorespiration, the synthesises of glycin from 2-phosphoglycolate