The full-length wild-type (FL WT) caspase-6 used in this study was derived from a synthetic, Escherichia coli codon-optimized (His)6 C-terminally tagged caspase-6 gene (Celtek Bioscience) that was ligated into the NdeI/BamHI sites of the pET11a vector. Caspase-6 variants (C163S, Y198A, and C163S/Y198A) as well as the N-terminally (His)6 tagged FL caspase-6 were generated using Phusion site-directed mutagenesis (Thermo Scientific) in the FL WT caspase-6 construct. Fully cleaved and active caspase-6 was also used in the form of a constitutive two chain (CT), which was designed to independently express the large and small subunits of caspase-6 with the prodomain (residues 1–23) and linker (residues 180–193) removed.9 (link)
Protocol detail
Caspase-6 Variants and Constructs
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization :
Other organizations : University of Massachusetts Amherst
Top 5 similar protocols
Variable analysis
independent variables
- Caspase-6 variants (C163S, Y198A, and C163S/Y198A)
- N-terminally (His)6 tagged FL caspase-6
- Fully cleaved and active caspase-6 in the form of a constitutive two chain (CT)
- Not explicitly mentioned
- Full-length wild-type (FL WT) caspase-6 derived from a synthetic, Escherichia coli codon-optimized (His)6 C-terminally tagged caspase-6 gene
- Full-length wild-type (FL WT) caspase-6 used as a control
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
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!