HYL1 | Hyponastic leave phenotype ds-RNA binding protein
AS06 136 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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40 µg of total protein from Arabidopsis thaliana rosette leaves extracted with extraction buffer (100 mM Tris HCl, pH 7.5; 10% glycerol; 5 mM EDTA; 5 mM EGTA; 150 mM NaCl; 0.75% Triton X-100; 0.05% SDS; 1 mM DTT; 1x Complete Mini EDTA-free protease inhibitor (Roche)) were separated on 12 % SDS-PAGE using semi-dry transfer and blotted 1h to PVDF. Blots were blocked with 5% milk in TBS-T O/N at 4°C with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 1,5 h at RT with 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 secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera, AS09 602) diluted to 1:50 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with chemiluminescence detection reagent, according to the manufacturer's instructions. Exposure time was 60 seconds. There were no other bands present on this blot in applied conditions
Courtesy of Dr. Dorota Raczyńska, M.Sc Agata Stepień Adam Mickiewicz University, Poland
HYL1 is nuclear localized, double stranded RNA-binding protein involved in microRNA (miRNA) biosynthesis. Synonyme: F21M12.9 protein EMBL AAB60726.1
Wang et al. (2019). The PROTEIN PHOSPHATASE4 Complex Promotes Transcription and Processing of Primary microRNAs in Arabidopsis. Plant Cell. 2019 Feb;31(2):486-501. doi: 10.1105/tpc.18.00556. (immunoprecipitation)
Su et al. (2017). The Protein Phosphatase 4 and SMEK1 Complex Dephosphorylates HYL1 to Promote miRNA Biogenesis by Antagonizing the MAPK Cascade in Arabidopsis. Dev Cell. 2017 Jun 5;41(5):527-539.e5. doi: 10.1016/j.devcel.2017.05.008.
Li et al. (2016). Intron Lariat RNA Inhibits MicroRNA Biogenesis by Sequestering the Dicing Complex in Arabidopsis. PLoS Genet. 2016 Nov 21;12(11):e1006422. doi: 10.1371/journal.pgen.1006422. eCollection 2016.
Francisco-Mangilet et al. (2015). THO2, core member of the THO/TREX complex, is required for micro RNA production in Arabidopsis. Plant J. 2015 May 14. doi: 10.1111/tpj.12874.
Raczynska et al. (2013). TheSERRATEprotein isinvolved inalternativesplicing inArabidopsis thaliana. Nucleic Acids Res. Oct 16.
Manavella et al. (2012). Fast-Forward Genetics Identifies Plant CPL Phosphatases as Regulators of miRNA Processing Factor HYL1. Cell Nov 9.
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