CERK1 | Chitin elicitor receptor kinase 1
AS16 4037 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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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
Wang et al. (2021) Arabidopsis PUB2 and PUB4 connect signaling components of pattern-triggered immunity. New Phytol. 2021 Dec 17. doi: 10.1111/nph.17922. Epub ahead of print. PMID: 34918346
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