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DCL1 | Dicer-like protein 1

265 €
Buy 2 items of this product for 198 €/each
Buy 3 items of this product for 180 €/each

AS12 2102  |  Clonality: Polyclonal  |  Host: Rabbit   |  Reactivity: Arabidopsis thaliana

PRODUCT INFORMATION IN PDF

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AS12 2102

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product information
Background

DCL1 (EC=3.1.26) is a ribonuclease (RNase) III involved in RNA-mediated post-transcriptional gene silencing (PTGS). Functions in the microRNAs (miRNAs) biogenesis pathway by cleaving primary miRNAs (pri-miRNAs) and precursor miRNAs (pre-miRNAs).
Synonymes: Protein ABNORMAL SUSPENSOR 1, Protein CARPEL FACTORY, Protein SHORT, INTEGUMENTS 1, Protein SUSPENSOR 1.

Immunogen

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana DCL1 sequence, Q9SP32 , At1g01040

Host Rabbit
Clonality Polyclonal
Clone
Purity
Affinity purified serum
Format Lyophilized
Quantity 100 µg
Reconstitution
For reconstitution add 50 µl of sterile water.
Storage

Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

Tested applications RIP (RNA immunoprecipitation assay), Western Blot (WB)
Related products
AS15 3100 | anti-DCL2 | Dicer-like protein 2, rabbit antibodies
AS12 2103
| anti-DCL3 | Dicer-like protein 3, rabbit antibodies
AS15 3105 | anti-DCL4 | Dicer-like protein 4
Additional information

Freezing and re-using of diluted anti-DCL1 antibodies is not recommended and will contribute to non-conclusive results.

application information
Recommended dilution

1 : 1000 with standard ECL (WB)

Expected | apparent MW

213 kDa

Confirmed reactivity

Arabidopsis thaliana

Predicted reactivity Arabidopsis thaliana
Not reactive in

Zea mays

Additional information

Recommend protein load is from 40-50 µg/well (using 1.5 mm spacers helps to obtain wider wells)

Selected references Yoshikawa (2016). Pre-microRNA processing activity in nuclear extracts from Arabidopsis suspension cells. J Plant Res. 2016 Nov 24. [Epub ahead of print]
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.

Application example
western blot using anti-DCL1 antibodies
Proteins from nuclear fraction and cytoplasmic fraction of Arabdiospsis thaliana were separated on  7 % SDS-PAGE using semi-dry transfer and blotted 1h to PVDF. Blots were blocked 2,5% milk in TBS-T  for over night at 4 degrees with agitation. Blot was incubated in the primary antibody (1:500; anti-DCL1, AS12 2102, lot 1508 (N) )) in PBS-T for 1h at RT with agitation. The antibody solution was decanted and the blot was washed 3 times for 15 min in PBS-T at RT with agitation. Blot was incubated in secondary antibody ( 1:10000; anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera cat no. AS09 607) for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturer's instructions. Exposure time was 360 seconds

Nuclear fraction: Nuclear fraction preparation was described in article: „The SERRATE protein is involved in alternative splicing in Arabidopsis thaliana” - Raczyńska et al., 2014.

Courtesy of M. Sc. Mateusz Bajczyk, Institute of Molecular Biology and Biotechnology, UAM, Poland.


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