活性部位反応型ＤＵＢプローブ

活性部位反応型ＤＵＢプローブ
脱ユビキチン化酵素（DUB)用の活性部位反応型プローブは、多くのDUBの活性部位に存在するシス
テイン残基に反応するよう設計されており、ユビキチンのC末端にDUB阻害剤であるVME（vinylmethyl ester）またはPA（propagylamide）が結合しています。（Figure 1）
UbiQ社ではVME（vinylmethyl ester）とPA（propagylamide）の2種類の反応基と、各反応基
で4種類のN末端タグ（TAMRA、Cy5、biotin、HA---＞Table 3）を付加したプローブ(Table 1)
をご用意しております。
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
VME1
PA2
tag
code
name
code
name
−
UbiQ-005
Ub-VME
UbiQ-057
Ub-PA
HA
UbiQ-035
HA-Ahx-Ahx-Ub-VME
UbiQ-078
HA-Ahx-Ahx-Ub-PA
Biotin
UbiQ-054
Biotin-Ahx-Ub-VME
UbiQ-076
Biotin -Ahx -Ub-PA
TAMRA
UbiQ-050
TAMRA-Ub-VME
UbiQ-058
TAMRA-Ub-PA
Cy5
UbiQ-071
Cy5-Ub-VME
UbiQ-072
Cy5-Ub-PA
UbiQ-L02
DUB probe explorer panel (our full range of all 10 DUB activity probes)
Table 2 – specifications of VME and PA warhead
VME
PA
targets USP and UCH DUBs
targets OTU, USP and UCH DUBs *
irreversible DUB inhibitor
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
Cy51a
Biotin1
HA1
fluorescence detection
fluorescence detection
affinity tag
affinity tag
• fast (in-gel) -, sensitive -,
• fast (in-gel) -, sensitive -,
• allows pull-down with
and distinct read-out.
• no background labelling
by cross reactivity as seen
in immunoblots
• exc 625 nm, abs 670 nm
• allows pull-down with HA
biotin binding tools
binding tools
• strongest known non-co- • influenza epitope tag
valent interaction
• detection by western-blot• detection by western-blotting
ting
fact sheet, October 2014
and distinct read-out.
• no background labelling
by cross reactivity as seen
in immunoblots
• exc 550 nm, abs 590 nm
literature
1(a) de Jong et al. ChemBioChem 2012, 13, 2251. (b) Altun et al. Chem Biol 2011, 18, 1401.
2
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.
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