活性部位反応型DUBプローブ (ACTIVITY-BASED DUB PROBES) 脱ユビキチン化酵素(DUB)用の活性部位反応型プローブは、ほとんどのDUBに存在する活性部位であるシステイン残基に反 応するよう設計されたC-末端のヘッド部分を持つユビキチンを基にしています。(Figure 1)UbiQ社はVME(vinylmethyl ester) とPA(propagylamide)の2タイプのヘッド部分を持つプローブとそれぞれに4種類の異なるN- 末端タグ(TAMRA、Cy5、biotin、 HA--->Table 3)を付加したプローブ(Table 1)をUbiQProbeとして販売しています。 Figure 1 western blotting, affinity tag direct in-gel read-out HA Biotin TAMRA Cy5 Ub HA Biotin TAMRA Cy5 Ub VME DUB PA DUB Table 1 - our range of DUB activity probes PA2 VME1 Tag Code Name 50 ug Code Name 50 ug − UbiQ-005 Ub-VME ご照会 UbiQ-057 Ub-PA ご照会 HA UbiQ-035 HA-Ahx-Ahx-Ub-VME ご照会 UbiQ-078 HA-Ahx-Ahx-Ub-PA ご照会 Biotin UbiQ-054 Biotin-Ahx-Ub-VME ご照会 UbiQ-076 Biotin -Ahx -Ub-PA ご照会 TAMR A UbiQ-050 TAMRA-Ub-VME ご照会 UbiQ-058 TAMRA-Ub-PA ご照会 Cy5 UbiQ-071 Cy5-Ub-VME ご照会 UbiQ-072 Cy5-Ub-PA ご照会 Table 2 – specifications of VME and PA warhead VME targets USP and UCH DUBs irreversible DUB inhibitor PA targets OTU, USP and UCH DUBs * irreversible DUB inhibitor which forms a covalent linkage that can be cleaved by acid treatment * According to a proteomics study the PA probes also target DUBs of the Machado-Josephin Domain family. 2 Table 3 – specifications of N-terminal tags TAMRA1a fluorescence detection fast (in-gel) -, sensitive -, and distinct read-out. no background labelling by cross reactivity as seen in immunoblots exc 550 nm, abs 590 nm Cy51a fluorescence detection fast (in-gel) -, sensitive -, and distinct read-out. no background labelling by cross reactivity as seen in immunoblots exc 625 nm, abs 670 nm Biotin1 affinity tag allows pull-down with biotin binding tools strongest known noncovalent interaction detection by westernblotting HA1 affinity tag allows pull-down with HA binding tools influenza epitope tag detection by westernblotting Literature 1. (a) de Jong et al. ChemBioChem 2012, 13, 2251. (b) Altun et al. Chem Biol 2011, 18, 1401. 2. (a) Ekkebus et al. J Am Chem Soc 2013, 135, 2867. (b) Sommer et al. Bioorg Med Chem 2013, 21, 2511. for a complete list of references we refer to the product group overview document. fact sheet, June 2014 〒541-0047 大阪市中央区淡路町 2 丁目 2 番 5 号 TEL:06-6231-6146 FAX:06-6231-6149 〒300-2655 茨城県つくば市万博公園西 F26 街区 1 インプレス 103 TEL:029-848-3220 FAX:029-848-3221
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