CGL78 | YCF54
AS10 936 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, C. reinhardtii, Synechocystis sp. PCC6803
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15 µg of total protein from Arabidopsis thaliana (ecotype Col-0), Chlamydomonas reinhardtii (strain 4A+) and Synechocystis sp. (strain PCC6803 / Kazusa), extracted with 56 mM Na2CO3, 56 mM DTT, 1 % (w/v) SDS, 12 % (w/v) Sucrose, 2 mM EDTA) were separated on 15% SDS-PAGE and blotted 1h to nitrocellulose membrane. Blot was blocked with 2% milk powder in TBS-T for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1:1000 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 TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:20 000 in 1% milk powder in TBS for 1h at RT with agitation. The blot was washed as above and developed for 5 min with chemiluminescent detection reagent according to the manufacturers instructions. Exposure time was 3 minutes.
Courtesy of Dr. Annabel Salinas Hartwig, Humboldt University, Germany
Currently this antibody has not been confirmed to detect CGL78 protein in Arabidopsis thaliana. If you are interested to use this antibody in Arabidopsis, please, contact us.
Please, omitt SDS from transfer buffer and reduce transfer time to 45 min. Nitrocellulose membrane is recommended and SDS is omitted to allow this LMW protein to bind tighter to the membrane.
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