High Sensitivity Streptavidin-HRP
Company: Thermo Fisher Scientific
Catalog#: 21130
Product evaluation:
Non-radioactive in vitro PINK1 Kinase Assays Using Ubiquitin or Parkin as Substrate
Author:
Date:
2016-10-05
[Abstract]  This protocol describes the in vitro phosphorylation of ubiquitin and Parkin by the kinase PINK1 using recombinant proteins. Both substrates, ubiquitin and Parkin, are phosphorylated at the conserved serine 65 residue (pS65-ubiquitin and pS65-Parkin). The protocol also includes the use of monomeric and K48- and K63-linked poly-ubiquitin chains as alternative substrates. Although there are commercially available antibodies, we have not tested their performance in this assay since, but ...

In vitro Detection of S-acylation on Recombinant Proteins via the Biotin-Switch Technique
Author:
Date:
2014-11-20
[Abstract]  Protein palmitoylation is the post-translational modification of proteins via the attachment of palmitate through acyl linkages. The nucleophile sulfhydryl group of cysteines is the common palmitoylation site. Covalent attachment of palmitate occurs on numerous proteins and is usually associated with directing protein localization to the endomembrane system. Detection of protein palmitoylation by in vivo labeling with tritium-labeled palmitic acid typically requires an autoradiographic ...

Binding to Secreted Bone Matrix in vitro
Author:
Date:
2014-02-20
[Abstract]  This method examines the bone matrix binding capacity of proteins. Using osteogenic differentiation medium, multipotent stromal cells (MSC) are induced to differentiate into osteocytes in vitro and to secrete bone matrix. The latter is confirmed using Alizarin red S staining, which detects the presence of calcific deposits (hydroxyapatite). These calcific deposits are used to test the bone binding properties of proteins. The binding to the calcific deposits is assessed by Western blot ...

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