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 Á¦Ç°¾È³»  Ç×ü,Ç׿ø,ŶƮ   Cancer Research  Epigenetics

SNAP-ChIP¢â K-MetStat panel

SNAP-ChIP (Sample Normalization & Antibody Profiling for ChIP) K-MetStat panelÀº lysine¿¡ methylationµÈ ´Ù¼öÀÇ dNucs ÆÐ³ÎÀ» ChIP ½ÇÇè¿¡ ÀÌ¿ëÇÔÀ¸·Î½á, Ç×üÀÇ Æ¯À̼ºÀ̳ª È¿À²¼ºÀ» Á¤È®È÷ ÆÇµ¶Çϵµ·Ï µµ¿ÍÁÝ´Ï´Ù.

EpiCypher
Á¦Ç° ¹®ÀÇÇϱâ
ÀμâÇϱâ

Ư¡
  • 3'¸»´ÜÀÇ Æ¯ÀÌÇÑ DNA ½ÃÄö½º("barcode")¿¡ ÀÇÇØ ½Äº°µÉ ¼ö Àִ ¼­·Î ´Ù¸¥ nucleosome 15°¡Áö¿Í 1°¡Áö unmodified nucleosomeÀÇ mixture poolÀÔ´Ï´Ù.
  • ChIP½ÇÇè½Ã, ¸Þƿȭ »óÅÂÀÇ ±¸Á¶Àû À¯»ç¼ºÀ¸·Î ÀÎÇÑ antibody cross-reactivity ¿©ºÎ È®ÀÎÀÌ Áß¿äÇÕ´Ï´Ù. SNAP-ChIP ÆÐ³ÎÀº ¸ðµç ChIP ½ÇÇè¿¡¼­antibody specificity¸¦ Á¤·®È­ ÇÏ¿© ÀÌ·± ¿À·ù¹®Á¦¸¦ ±Øº¹ÇÕ´Ï´Ù.
  • ¸ðµç ChIP work flow¿¡ ÀÌ¿ëÇÒ ¼ö ÀÖ½À´Ï´Ù.
  • PanelÀÇ Modification site´Â ¸ðµÎ lysine methylation¿¡ ÀÇÇØ °áÁ¤µÇ±â ¶§¹®¿¡, À¯¿¬¼ºÀ» ±Ø´ëÈ­ÇÏ°í »ó¼¼ÇÑ Ç×ü ±³Â÷ ¹ÝÀÀ µ¥ÀÌÅÍ È®ÀÎÀÌ °¡´ÉÇÕ´Ï´Ù.
  • ³·Àº lot-to-lot variation À» Á¦°øÇÕ´Ï´Ù. (³ôÀº ÀçÇö¼º)
  • Enzyme screening assays ¹× Effector protein binding experiments ½ÇÇè¿¡¼­ ChIP spike-in ´ëÁ¶±ºÀ¸·Î ÀûÇÕÇÕ´Ï´Ù.
  • Reference: SNAP-ChIP is adapted from Grzybowski AT et al (2015) Mol Cell 58: 886-889.



  • Composition:1 unmodified mononucleous + 15 histone H3 or H4 posttranslational modifications (PTMs, created by a proprietary semi-synthetic method): H3K4me1, H3K4me2, H3K4me3, H3K9me1, H3K9me2, H3K9me3, H3K27me1, H3K27me2, H3K27me3, H3K36me1, H3K36me2, H3K36me3, H4K20me1, H4K20me2 and H4K20me3
  • Formulation: Purified recombinant mononucleosomes, containing a mixture of 16 (1 unmodified plus 15 unique) H3 and H4 PTMs in 10 mM sodium cacodylate, pH 7.5, 100 mM NaCl, 1 mM EDTA, 50% glycerol (w/v), 1x Protease Inhibitor cocktail, 100 ug/mL BSA, 10 mM β-mercaptoethanol. Molarity = 0.6 nM. MW = ~199382.1 Da (average MW of all 16 nucleosomes).

 

 Fig 1.  K-MetStat PanelÀ» ±¸¼ºÇÏ´Â 16°³ÀÇ dNucs Á¶ÇÕ ¹æ¹ýÀº ´ÙÀ½°ú °°½À´Ï´Ù. Modified dNucs
 ÀÇ °æ¿ì, H3K4, H3K9, H3K27, H3K36, H4K20¿¡ lysine methylation(me1, me2, me3)ÀÌ µÇ¾î ÀÖ°í,
 °¢°¢ ƯÁ¤ÇÑ ¼­¿­·Î DNA barcode µÇ¾î ÀÖ½À´Ï´Ù. ÆÐ³ÎÀº ÀÌ 15°³ÀÇ modified¿Í 1°³ÀÇ unmodified
 ·Î ±¸¼ºµË´Ï´Ù.





 Fig 2.  ƯÁ¤ DNA ½ÃÄö½º(barcode)·Î ½Äº°µÇ´Â PTMÀ» °¡Áø ÀçÁ¶ÇÕ dNucs poolÀº IPÀü¿¡ sample  
 chromatin¿¡ ÷°¡µÇ°í, barcoded nucleosomeÀ» captureÇÔÀ¸·Î½á Ç×ü ƯÀ̼ºÀ» ¸ð´ÏÅ͸µ  ÇÒ¼ö
 ÀÖ½À´Ï´Ù.