
Overview:
SETD8 or SET domain containing lysine methyltransferase 8 is a lysine methyltransferase that monomethylates p53 in human cell lines. The monomethylation by SETD8 suppresses p53-mediated transcriptional activation of highly responsive target genes, such as p21 (CDKN1A) and PUMA (BBC3) but it has little influence on weak p53 targets (1). Depletion of SET8 augmented the proapoptotic and checkpoint activation functions of p53, and SETD8 expression is downregulated during DNA damage. SETD8 is a Wnt signaling mediator and is recruited by LEF1/TCF4 to regulate the transcription of Wnt-activated genes, possibly through H4K20 monomethylation at the target gene promoters (2).
Gene Aliases:
PR-Set7, SET07, SET8
Genbank Number:
BC050346
References:
1. Shi, X. et al: Modulation of p53 function by SET8-mediated methylation at lysine 382. Molec. Cell 27: 636-646, 2007. 2. Li, Z. et al: Histone H4 Lys 20 monomethylation by histone methylase SET8 mediates Wnt target gene activation. Proc Natl Acad Sci U S A. 2011 Feb 22;108(8):3116-23.
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植物材料 用最精确的方法,称取不超过100mg的植物材料,置于处理过的研钵,加入液氮进行研磨 将研磨得到的粉末,快速转移至无RNase,并经过液氮冷却的2mL离心管(自备),
但是,这样做的前提的,RNA提得要好,浓度要高,这样免得RNA降解到不能用了。因为在有二价阳离子的(DNAseI buffer含有)情况下,RNA在60度以上时必然发生非酶促降解。
想搞清一个问题,如何区别是因为small RNA 引起的调控呢?还是tRNA引起的调控,有没有有效的分离这两种RNA的办法呢?
样品处理:
a. 植物组织:取新鲜或-70℃冻存100mg组织在液氮中研磨,把粉末加入到1ml裂解液中混匀。
b. 动物组织:取新鲜或-70℃冻存100mg组织加1ml裂解液,用组织研磨杵或匀浆器匀浆处理。
c. 贴壁细胞:直接在培养板中加入裂解液裂解细胞,每106细胞加1ml 裂解液。用取样器吹打混匀。

