Anti-Lhcb1 | LHCII type I chlorophyll a/b-binding protein

Pierrick
This antibody gives a clear single band with Arabidopsis leaf protein extracts (from leaf punch). I blocked the membranes with TBST + 3% milk for 1h at RT. The primary antibodies were diluted in TBST + 3% milk 1:5000 and incubated with the membranes for 1 h at RT with agitation. I´m reusing the diluted antibodies for up to 5 to 10 western blots without any significant loss of signal (Diluted antibodies are stored at -20°C). The secondary antibodies were diluted in TBST + 3% milk 1:10000 and incubated with the membranes for 1 h at RT with agitation.	2020-11-16 @ 10:06:52
Tanja Göbel
I´m using the antibodies for the immunodetection on western blots with the fluorescent based licor odyssey system (with the IRDye secondary antibodies Donkey anti rabbit IgG 800CW and 680 (Licor)). I´m analysing whole protein or chloroplasts protein extracts optained from Arabidopsis thaliana. Proteins were either precipitated with TCA/Aceton or directly solubilized with NP-40 in a tris-buffer. The only problem with the unprecipitated samples is that one gets a very strong backround in the 680nm channel due to the chlorophyll-fluorescence. The primary antibodies were diluted in TBS with 0,02% Sodiumazid according to the recommanded dilution (for ECL) in the data sheet and the membranes were incubated for 1 h at RT with agitation. The membranes are blocked with BSA in TBS and washed with TBS-T. I´m reusing the diluted antibodies for up to 5 to 10 western blots without any significant loss of signal (diuluted antibodies are stored at 4°C). The signals I´m getting with the antibodies are very clear and hieghly specific, with only low (anti-Lhcb1 and 2) or nearly no (anti-RbcL and RA) unspecific binding. Usually, I´m loading 10 µg protein and seperate them on 10% Mini-gels. So far I had no problems with Agrisera-antibodies.	2012-09-25 @ 13:13:52