immunoprecipitation (IP): 5-10 μg using cell lystes of HEK-293T indirect immunofluorescence: 2-5 μg/mL using paraformaldehyde-fixed NIH-3T3 cells over-expressing human DCP1A or using paraformaldehyde-fixed HEPG2 cells western blot: 1-2 μg/mL using cell lysates of HEK-293T
Dcp1 colocalizes with Dcp2 in distinct cytoplasmic foci along with other proteins involved in the 5′ to 3′ mRNA decay. These foci are termed PB (processing bodies) or DCP-bodies. Anti-DCP1A (C-terminal) is produced in rabbit using as immunogen a synthetic peptide corresponding to a sequence at C-terminal of human DCP1A conjugated to KLH. Two distinct genes of human DCP1 were identified, DCP1A and DCP1B, which share ~70% homology in their N-terminal and ~30% homology in their full length.
應用
Anti-DCP1A antibody produced in rabbit is suitable for immunoprecipitation at a working concentration of 5-10 μg using cell lystes of HEK-293T, indirect immunofluorescence at 2-5 μg/mL using paraformaldehyde-fixed NIH-3T3 cells over-expressing human DCP1A or using paraformaldehyde-fixed HEPG2 cells and western blot analysis at 1-2 μg/mL working concentration using cell lysates of HEK-293T. It was used as a primary antibody at a working dilution of 1:200 in the immunofluorescence experiment of HeLa cells treated with 5-fluorouracil to study the assembly of stress granules based on RNA incorporation.
Dcp1 cleaves the m7G mRNA cap in the 5′ to 3′ mRNA decay pathway, in association with Dcp2 and Hedls complex. Decapping is a critical and highly regulated step in the turnover of mRNA which involves decapping enzymes that hydrolyze the cap structure at the 5′ mRNA.
外觀
Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide.
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