Microscopic detection of quenched activity-based optical imaging probes using an antibody detection system: localizing protease activity.
Microscopic detection of quenched activity-based optical imaging probes using an antibody detection system: localizing protease activity.
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DOI：
10.1007/s11307-014-0736-1
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发表时间：
2014-10
影响因子：
3.1
通讯作者：
Basilion, James P.
中科院分区：
文献类型：
--
作者：
Walker, Ethan;Gopalakrishnan, Ramamurthy;Bogyo, Matthew;Basilion, James P.
关键词：
The cysteine cathepsins are a family of proteases that play important physiological roles in both normal cellular physiology and many human diseases. In cancer, the activity of many of the cysteine cathepsins is up-regulated and can be exploited for tumor imaging both in vivo and ex vivo. To characterize the behavior of a topically applied quenched fluorescent activity-based probe, GB119, ex vivo, we developed a basic immunohistochemistry technique to identify unquenched GB119 within tissue. Immunoblot assays were used to validate the utility of an anit-Cy5 antibody for the detection of unquenched GB119 generated by cathepsin-L. Following validation the anti-Cy5 antibody an immunohistochemical procedure was developed to detect the presence of unquenched GB119 in frozen sections of brain tumors derived from an orthotopic mouse model. These studies demonstrate that the anti-Cy5 antibody preferentially recognizes unquenched GB119 and that this differential can be used to identify the regions within the brain and the tumor that contained unquenched GB119. Using H&E staining and antibodies against other biochemical markers it was further determined that unquenched GB119 was localized to the peri-tumor space and co-localized with cathepsin-L expression. Our data indicate that this methodology allows high-resolution detection of unquenched GB119 that can be correlated with other immunohistological stains.
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影响因子：
--
作者：
Verdoes M;Edgington LE;Scheeren FA;Leyva M;Blum G;Weiskopf K;Bachmann MH;Ellman JA;Bogyo M
通讯作者：
Bogyo M
影响因子：
3.7
作者：
Blum G;Weimer RM;Edgington LE;Adams W;Bogyo M
通讯作者：
Bogyo M
DOI：
10.1007/978-3-642-16861-1_2
发表时间：
2011-01-01
期刊：
EXTRACELLULAR MATRIX DEGRADATION
影响因子：
--
作者：
Broemme, Dieter;Wilson, Susan
通讯作者：
Wilson, Susan
影响因子：
14.8
作者：
Blum, Galia;von Degenfeld, Georges;Bogyo, Matthew
通讯作者：
Bogyo, Matthew
影响因子：
3.7
作者：
Cutter, Jennifer L.;Cohen, Nathan T.;Basilion, James P.
通讯作者：
Basilion, James P.