One μg of total protein from sample homogenates was used. The reaction mixture contained 50 mM Tris–HCl pH = 8, 1 mM DTT, and 40 μM Suc-Leu-Leu-Val-Tyr-AMC (Sigma Aldrich) substrate for chymotrypsin-like activity or 40 μM of Boc-Leu-Ser-Thr-Arg-AMC for trypsin-like activity or 100 μM of Ac-Gly-Pro-Leu-Asp-AMC (Sigma Aldrich) for Caspase-like activity. To distinguish the 26S proteasome activity from the 20S proteasome activity, 5 mM ATP was added to the reaction mixture. The mixture was then incubated for 30 min at 37 °C. The reaction was stopped by adding 100 mM monochloroacetate and 30 mM sodium acetate pH = 4.3. Fluorescence was determined by measuring the release of AMC (λ excitation: 370 nm, λ emission: 410 nm), using a Perkin Elmer LS 30 spectrofluorometer.
Ac gly pro leu asp amc
Manufactured by Merck Group
Ac-Gly-Pro-Leu-Asp-AMC is a synthetic peptide substrate used for the detection and quantification of protease enzyme activity in research applications. It consists of the amino acid sequence Ac-Gly-Pro-Leu-Asp, with an AMC (7-amino-4-methylcoumarin) fluorogenic reporter group attached to the C-terminus. When cleaved by a protease, the AMC group is released, resulting in a fluorescent signal that can be measured.
Automatically generated - may contain errors
2 protocols using ac gly pro leu asp amc
1
Proteasome Activity Assay
2
Proteasome Activity Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
One μg of total protein from sample homogenates was used. The reaction mixture contained 50 mM Tris–HCl pH = 8, 1 mM DTT, and 40 μM Suc-Leu-Leu-Val-Tyr-AMC (Sigma Aldrich) substrate for chymotrypsin-like activity or 40 μM of Boc-Leu-Ser-Thr-Arg-AMC for trypsin-like activity or 100 μM of Ac-Gly-Pro-Leu-Asp-AMC (Sigma Aldrich) for Caspase-like activity. 5 mM ATP was added to the reaction mixture to distinguish 26S proteasome activity from 20S proteasome activity. The mixture was then incubated for 30 min at 37 °C, and stopped by adding 100 μM monochloroacetate and 30 mM sodium acetate pH = 4.3. Fluorescence was determined by measuring the release of AMC (λ excitation: 370 nm, λ emission: 410 nm), using a Perkin Elmer LS 30 spectrofluorometer.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!