| 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 | | antibody format | | rabbit | polyclonal | | affinity purified serum | lyophilized | | | quantity | | 200 µg | for reconstitution add 200µl, 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 | | western blot (WB) | | related products | | AS12 2103 | DCL3 | Dicer-like protein 3 | | additional information | | to be added when available | |
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 | | no confirmed exceptions from predicted reactivity are currently known |
| additional information | | Recommend protein load is from 40-50µg/well (using 1.5 mm spacers helps to obtain wider wells) |
| selected references | | to be added when available, antibody released in August 2012. |
application example
50 µg of total protein from Arabidopsis thaliana total cell extract which came from the ground, frozen powder and has been directly transferred to 2x Laemmli sample buffer was separated on 15 % SDS-PAGE and blotted over night at 64 mA to nitrocellulose (wet transfer). Blots were blocked with Roti-block over night at 4°C agitation. Blot was incubated in the primary antibody at a dilution of 1: 2000 (1) and 1: 500 (2) 1h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in PBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:10 000 for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturers instructions. Exposure time was 1 min.
Courtesy of Dr. Sascha Laubinger, ZMBP, Germany
bio-equip.com
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