UbiQ-Probesについて

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
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