HYL1 | Hyponastic leave phenotype ds-RNA binding protein
AS06 136 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

Data sheet | Product citations | Protocols | Customer reviews |
Product Information
Immunogen
KLH-conjugated synthetic peptide derived from Arabidopsis thaliana Hyl1 protein sequence UniProt: O04492, TAIR: At1g09700
Host
Rabbit
Clonality
Polyclonal
Purity
Serum
Format
Lyophilized
Quantity
50 ĩl
Reconstitution
For reconstitution add 50 ĩl of sterile water
Storage
Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
Tested applications
RNA Immunoprecipitation (RIP), Western blot (WB)
Recommended dilution
1: 1000 (WB)
Expected | apparent MW
45,5 kDa | 68-70 kDa
Reactivity
Confirmed reactivity
Arabidopsis thaliana
Predicted reactivity
Arabidopsis thaliana
Not reactive in
No confirmed exceptions from predicted reactivity are currently known
Application examples
Application examples
Courtesy of Dr. Dorota Raczyńska, M.Sc Agata Stepień Adam Mickiewicz University, Poland
Application example

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

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
Additional information
Background
Background
HYL1 is nuclear localized, double stranded RNA-binding protein involved in microRNA (miRNA) biosynthesis. Synonyme: F21M12.9 protein EMBL AAB60726.1
Product citations
Selected references
Ren et al. (2020). BcpLH organizes a specific subset of microRNAs to form a leafy head in Chinese cabbage (Brassica rapa ssp. pekinensis). Hortic Res. 2020 Jan 1;7:1. doi: 10.1038/s41438-019-0222-7.
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.
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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