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

Histone Peptide Arrays

EpiTriton Histone Peptide Array´Â effector protein, antibody, enzyme°£ÀÇ »óÈ£ÀÛ¿ëÀ» ½ºÅ©¸®´×Çϱâ À§ÇÑ Histone library array ÀÔ´Ï´Ù.

EpiCypher
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  • EpiTriton Histone Peptide Array´Â effector protein, antibody, enzyme°£ÀÇ »óÈ£ÀÛ¿ëÀ» ½ºÅ©¸®´×Çϱâ À§ÇÑ Histone library array ÀÔ´Ï´Ù.
  • ¿©·¯ Á¾·ùÀÇ PTM histone peptide spotÀ¸·Î ±¸¼ºµÇ¾î ÀÖ½À´Ï´Ù.
  • Çü±¤À̳ª HRP·Î ÄÚÆÃµÈ Ç×ü·Î °¡½ÃÈ­ ÇÑ ÈÄ Çü±¤½ºÄ³³Ê³ª ECL ½ºÄ³³Ê·Î È®ÀÎÇÕ´Ï´Ù.
  • ¸ðµç modified peptide´Â HPLC purificationÀ» °ÅÃÆ½À´Ï´Ù. (>95% pure, 20 amino acids)
  • ÇÑ ½½¶óÀ̵å À§¿¡ 3°³ÀÇ subarray·Î ±¸¼ºµÇ¾î 3°¡ÁöÀÇ ´Ù¸¥ Á¶°ÇÀ¸·Î ½ÇÇèÇÏ½Ç ¼ö ÀÖ½À´Ï´Ù. (multiwell array gasket ÀÌ¿ë)
  • °¢ peptide´Â 3°³¾¿ ³ª¶õÈ÷, 18¹ø¾¿ ¹Ýº¹ÇÏ¿© spotted µÇ¾î ÀÖ¾î, °á°úÀÇ ½Åºù¼ºÀ» º¸ÀåÇÕ´Ï´Ù.
  • ½ºÄµÇÑ °á°ú, peptide spot tracer´Â greenÀ¸·Î, Ç×ü¿¡ ´ëÇÑ ´Ü¹éÁúÀº red·Î °¡½ÃÈ­ µË´Ï´Ù. 3°³ÀÇ ¿¬¼ÓÀûÀÎ spotÀ» º¸ÀÌ´Â ºÎºÐÀÌ ÀÇ¹Ì ÀÖ´Â °á°úÀÔ´Ï´Ù.

 

  • EpiTitan™ Histone Peptide Array Design:
    Each EpiCypher EpiTitan™ Histone Peptide Array contains over 265 biotinylated histone peptides (20 amino acids in length or more) immobilized on a streptavidin-coated glass slide. Peptides are spotted as two identical sub-arrays, labeled A and B. Within each sub-array, each peptide is spotted twice in groups of 3 (red dots), for a total of 12 spots per peptide on each array.

  • EpiTitan™ Histone Peptide Array Workflow:
    For detection of the interaction of an effector protein with peptides on the array as shown above, you need a primary antibody to the protein (or to an affinity tag) and a fluorescently-labeled (or HRP-conjugated for ECL detection) secondary antibody to the primary. This is much like the detection procedure employed in immunofluorescence miscroscopy. For analysis of histone antibody specificity, you need a primary antibody to a histone modification and a labeled (fluorescent or HRP-conjugated) secondary antibody recognizing the primary antibody.


  • Examine the selectivity and specificity of histone modification antibodies
  • Analyze the specificity of histone binding proteins
  • Identify substrates for histone-modifying enzymes






  • Rothbart SB et al (2015). An Interactive Database for the Assessment of Histone Antibody Specificity. Mol Cell 59: 502-511. LINK
  • Ali M et al (2015). Molecular insight into inhibition of the methylated histone-plant homeodomain complexes by calixarenes. J Biol Chem. LINK
  • Zhang ZM et al (2015). An Allosteric Interaction Links USP7 to Deubiquitination and Chromatin Targeting of UHRF1. Cell Rep. LINK
  • Tong Q et al (2015). An acetyl-methyl switch drives a conformational change in p53. Structure 23: 322-331. LINK
  • Tong Q et al (2015). Structural plasticity of methyllysine recognition by the tandem tudor domain of 53BP1. Tong Q et al. 2015. Structure 23: 312-321. LINK
  • Rothbart SB et al (2015). An Interactive Database for the Assessment of Histone Antibody Specificity. Mol Cell 59: 502-511. Link
  • Greer EL et al (2014). A histone methylation network regulates transgenerational epigenetic memory in C. elegans. Cell Rep 7: 113-126. Link
  • Kim HS et al (2014). Identification of a BET family bromodomain/casein kinase II/TAF-containing complex as a regulator of mitotic condensin function. Cell Rep 6: 892-905. Link
  • Klein BJ et al (2014). The histone-H3K4-specific demethylase KDM5B binds to its substrate and product through distinct PHD fingers. Cell Rep 6: 325-335. Link
  • Kinkelin K et al (2013). Structures of RNA polymerase II complexes with Bye1, a chromatin-binding PHF3/DIDO homologue. PNAS USA 110: 15277-15282. Link
  • Ali M et al (2013). Molecular basis for chromatin binding and regulation of MLL5. PNAS USA 110: 11296-11301. Link
  • Rothbart SB et al (2013). Multivalent histone engagement by the linked tandem Tudor and PHD domains of UHRF1 is required for the epigenetic inheritance of DNA methylation. Genes Dev 27: 1288-1298. Link
  • Gatchalian J et al (2013). Dido3 PHD modulates cell differentiation and division. Cell Rep 4: 148-158. Link
  • Cai L et al (2013). An H3K36 Methylation-Engaging Tudor Motif of Polycomb-like Proteins Mediates PRC2 Complex Targeting. Mol Cell 49: 571-582. Link
  • Rothbart SB et al (2012). Association of UHRF1 with methylated H3K9 directs the maintenance of DNA methylation. Nat Struct Mol Biol 19: 1155-1160. Link
  • Rothbart SB, Krajewski K, Strahl BD, Fuchs, SM (2012). Peptide microarrays to interrogate the histone code. Methods Enzymol 512: 107-135. Link
  • Fuchs SM, Krajewski K, Baker RW, Miller VL, Strahl BD (2011). Influence of combinatorial histone modifications on antibody and effector protein recognition. Curr Biol 21: 53-58. Link