Storage Condition ： Store at 0-5oC
Modification of protein thiol is one of the most important post-translational modifications and it occurs depending on the redox state in cells. Protein S-nitrosylation is NO (nitric oxide)-dependent modification of protein thiols and is crucial for regulation of cellular functions such as transcription, protein expression, and signal transduction.
-SulfoBiotics- Protein S-Nitrosylation Monitoring Kit allows to detect S-nitrosylated proteins by gel-electrophoretic analysis. This kit contains chemical reagents for blocking of free thiols on proteins, reducing of S-nitrosylated thiols, and labeling of the reduced thiols. After blocking free thiols of protein, S-nitrosylated thiols are selectively reduced by the reducing agent, and labeled with Protein-SHifter Plus, which is a novel maleimidyl compound consisted of a high molecular weight. When one molecule of Protein-SHifter Plus binds to a thiol group of protein, a mobility shift corresponding to about 15 kDa of molecular mass is observed by the gel-electrophoretic analysis. Thus, the number of S-nitrosylated thiol group on a protein can be clearly identified by SDS-PAGE through the mobility shift assay. In addition, the Protein-SHifter Plus moiety can be cleaved from the labeled protein in a gel with UV irradiation after gel-electrophoresis because Protein-SHifter Plus has a UV photocleavable moiety in the molecule. Therefore, the protein treated with UV irradiation can be transferred from the gel to PVDF membrane and detected on the membrane similar to the unlabeled protein by a specific antibody.
Figure 1 Schematic Protocol of Protein S-Nitrosylation Monitoring Kit
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