LHCSR3

Application example



Follwoing samples: 0.1 µg of LhcSR3 IB + HisTag (1), 0.05 µg of LhcSR3 IB + HisTag (2), 5 µg of Chlamydomonas reinhardtii wild type (CC124) total protein extract of photoautotrophically grown cells in light intensity: 60 µE (3), 5 µg of Chlamydomonas reinhardtii wild type (CC124) total protein extract of photoautotrophically grown cells in high light intensity: 500 µE (4) were separated on 15% Tris-Glycine SDS PAGE and blotted overnight to PVDF using tank transfer. Blots were blocked with 5% BSA/milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 1h at RT 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 PBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera, AS09 602) diluted to 1:10 000 for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturer's instructions.