CRY2 | Cryptochrome 2

Samples:
50 µg of Arabidopsis thaliana whole seedlings (7 days old)
1 - cry1-304 (overexpressing line)
2 - cry2-1 (deletion mutant of both isoforms)
3 - cry1-304 cry2-1 (substitution mutant)

Mark: MW markers: PageRuler™ Plus Prestained Protein Ladder from Thermo Scientific™ (26619)

8 µl/well of total protein extracted freshly from 50 ug of Arabidopsis thaliana seedlings (7-day WL grown) with 8M Urea, 0.35M Tris-Cl (pH 7.5), 1× protease inhibitor cocktail. and denatured with 6x SDS buffer at 95°C for 10 min. Samples were separated o 4-12 % gradient SDS-PAGE (Invitrogen) and transferred 7 min to nitrocellulose (pore size of 0.2 µm), using transfer device (iBlot2, Thermofischer). Blot was blocked with 2 % milk 1h/RTwith agitation. Blot was incubated in the primary antibody at a dilution of 1: 1,000 for ON/4°C without agitation. The antibody solution was decanted and the blot was washed 3 times for 10 min in TBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (goat anti-rabbit HRP conjugated AS09 602) diluted to 1:25 000 in for 3h/RT with agitation. The blot was washed as above and developed with AgriseraECLSuperBright AS16 ECL-S-100, using Summit QCP. Exposure time was 5 seconds.

Courtesy of Dr. Sanghwa Lee, Salk Institute for Biological Studies, USA

Alternative protein names: Blue light photoreceptor1, Protein PHR homolog 11 AtPHH1, Protein SUPPRESSOR OF elf3 20