HEN1 | HUA ENHANCER 1
AS15 3095 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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50 µg of total protein from Arabidopsis thaliana whole vegetative rosette wild type Col-0 (a) and HEN1 overexpression mutant (b) extracted with extraction buffer (50 mM Tris pH 7.5; 150 mM NaCl; 1 mM EDTA; 10 % v/v Glycerin; 1 mM DTT, 1x Complete Protease Inhibitor Cocktail, Roche) and denatured with Laemmli buffer at 95°C/5 min., were separated on 7.5 % SDS-PAGE and blotted 1.5 h to PVDF using tank transfer. Blots were blocked with blocking buffer (5% milk powder; 1x TBS; 0.1% Tween-20) overnight at 4°C with agitation. Blot was incubated in the primary antibody at a dilution of 1:1000 for 2h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly and then washed tree times for 15 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 1:20 000 in blocking buffer for 1h at RT with agitation. The blot was washed as above and developed for 5 min with Amersham ECL Prime and expose to Amersham Hyperfilms ECL for 20 seconds.
Courtesy of Dr. Pablo Manavella, Instituto de Agrobiotecnología del Litoral (IAL), Argentina
HEN1 (HUA ENHANCER 1) is a methyltransferase that can methylate 3'-end of miRNAs, siRNas and trans-acting small interfering RNAs (ta-siRNAs). This protein protects the 3'-end of sRNAs from uridylation activity and degradation by adding a methyl group to the ribose of the last nucleotide of sRNAs.
Alternative protein names: Small RNA 2'-O-methyltransferase, Protein CORYMBOSA 2, S-adenosylmethionine-dependent RNA methyltransferase HEN1.
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