CPK-EF | Calcium-dependent protein kinase EF domain
AS11 1807 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, H. vulgare, Populus sp., T. aestivum
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20 µg of total protein from Hordeum vulgare extracted with isolation buffer with 6M urea were separated on 12% SDS-PAGE and blotted 1h to PVDF using semi-dry transfer. Blots were blocked with 5% milk in TBS-T 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 once, then washed 3 times for 10 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:25 000 in TBS-T containing2% milk 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. Exposure time was 300 seconds.
Courtesy of Dr. Lucyna Misztal, Institute of Molecular Biology and Biotechnology, Poznań, Poland
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