CERK1 | Chitin elicitor receptor kinase 1
AS16 4037 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
1: 1000 - 1: 3500 (WB)
Nicotiana benthamiana, Solanum lycopersicum
Total protein from 16-day old Arabidopsis thaliana Col-0 (wild-type) and cerk1-2 null mutant (Cao et al., 2014). Total protein was extracted using Buffer H (Heese et al., 2007; LaMontagne et al., 2016) except that 40 nM Calyculin was used. 70 µg of total protein was denatured at 65°C for 5 min, separated on an 8% SDS-PAGE and transferred for 1h using a tank transfer system to nitrocellulose membrane. Blots were blocked with 1x PBS (Fisher Scientific BP665-1) + 0.1 %Tween 20 (PBS-T) + 5% milk for 1.5h at room temperature (RT) with agitation. Blots were incubated with primary antibody at a dilution of 1: 1000 overnight at 4°C with agitation in 1x PBS-T + 5% milk. The antibody solution was decanted, and the blot washed four times (1x 5min and 3x 6min) in 1x PBS-T at RT with agitation. The blot was then incubated with secondary antibody (Goat anti Rabbit IgG (H&L)–HRP from Agrisera (AS09 602) diluted to 1:10,000 in 1x PBS-T + 5% milk for 4h at RT with agitation. The blot was washed as above and developed for 5 min with Amersham ECL (RPN2106). Exposure time was about 10 min.
Courtesy of Grant Mc Gowan, Gayani Ekanayke & Dr. Antje Heese, Division of Biochemistry, Interdisciplinary Plant Group (IPG), University of Missouri; Columbia, MO, USA.
- Cao, Y. et al. (2014). The kinase LYK5 is a major chitin receptor in Arabidopsis and forms a chitin-induced complex with related kinase CERK1. eLife 2014;3:e03766.
- Heese, A. et al. (2007). The receptor-like kinase SERK3/BAK1 is a central regulator of innate immunity in plants. PNAS. Vol. 104 no. 29 p. 12217–12222.
- LaMontagne et al., (2016). Isolation of Microsomal Membrane Proteins from Arabidopsis thaliana. Current Protocols in Plant Biology 1:1-18. doi: 10.1002/cppb.20020