EDS1 | Enhanced disease susceptibility 1
AS13 2751 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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71.6 kDa | 72 kDa
1 -40 ug of Arabidopsis thaliana whole leaf extract, non treated
2 -40 ug of Arabidopsis thaliana whole leaf extract, treated with PstAvrB 3h
3- 40 ug of Arabidopsis thaliana whole leaf extract, old-6 weeks
4-40 ug of Solanum lycopersicum (negative control) 8 discs extract, control leaf
5-40 ug of Solanum lycopersicum (negative control) 8 discs extract, wounded leaf
6-40 ug of Arabidopsis thaliana whole leaf extract, mutant eds1
7-40 ug of Arabidopsis thaliana whole leaf extract, mutant eds1
Mark: MW markers: 5uL Thermo scientific pageruler plus prestained ladder
40 ug/well of total protein was extracted freshly from Arabidopsis thaliana and Solanum lycopersicum (see above) with a buffer: Tris-HCl pH 7,5 50mM; 10% glycerol; 1mM EDTA; 200mM NaCl; 1mM DTT; 0,1% Triton X-100; Sigma protease cocktail; sigma phosphatase inhibitor cocktail; 140 mM pepstatine A and 0,1mM PMSF in MiliQ water (samples 1-5) and Tris-HCl 50mM pH 7.8; 0.2% triton x-100; Sigma protease inhibitor cocktail; 1% PMSF and 1.5% pepstatin A in MiliQ water; (samples 6-7) and denatured with SDS buffer (Tris-HCl pH 6,8, 8% SDS, 40% glycerol) and 1ul DTT 0,5 M at 95°C for 5 min. Proteins were separated on 12 % SDS-PAGE and blotted 1h to PVDF (pore size of 0,45 µm), using semi-dry. Blot was blocked with 3 % non-fat milk for 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 with agitation in 3% milk-TBS-T 0,1 %T ON/4°C. The antibody solution was decanted and the blot then washed three times for 10 min 3% milk-TBS-0,1%T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated AS09 602) diluted to 1:10 000 in 3% milk-TBS-0,1%T for 1h/RT with agitation. The blot was washed three times wit TBS and developed for 4 min (300uL + 300 uL / membrane) with either Agrisera ECLSuperBright. Exposure time was 1800 seconds in total. The image correspond to 1660-second exposure.
Courtesy of Dr. María C. Romero-Puertas, Department of Biochemistry, Cell and Molecular Biology of Plants, CSIC, Spain
EDS1 (Enhanced disease susceptibility 1) is a protein with lipase activity involved in aerencyma formation, lipid metabolic process, response to hypoxia and systemic acquired resistance. Alternative names: Putative disease resistance protein EDS1, Putative uncharacterized protein T17F15.40.
Chakraborty et al. (2018). Epigenetic and transcriptional control of chickpea WRKY40 promoter activity under Fusarium stress and its heterologous expression in Arabidopsis leads to enhanced resistance against bacterial pathogen. Plant Science, doi.org/10.1016/j.plantsci.2018.07.014
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