Nascent Protein Synthesis

Nascent protein synthesis tools

Several options are available depending on the needs of the experiment. These modified amino acids offer options to be incorporated into proteins during synthesis, chemical modification or use of the expanded genetic code to incorporate modified amino acids with high specificity & translational fidelity. (1) Detection levels can be in the low femtomole range in 1D Western blots, and some can be used in LC-MS/MS and MADLI   MS analysis.

The site specific labelling can be particularly useful when specific functionality of the protein is required downstream, enabling orientation of the protein, or control of the level of conjugation.

Products : -

O-propargyl-puromycin (OPP) incorporates an alkyne terminal group during protein synthesis and label with a CUAAC labelling reaction. Terminates protein synthesis, but is not an amino acid analogue, so can be added to cells in complete growth media. San be used for mammalian cells and to assay nascent proteins in whole organisms. Detect with an azide reagent.

Click & Go OPP Kits - Contains OPP labelling reagent and detection reagents. Cells can be independently or co-incubated with the drug & OPP, fixed & permeabilized and the copper click reaction performed with the included fluorescent dye for visualization. Each kit is enough for imaging 25 coverslips or 250 wells (based on 96 well plate).

4-Azido-L-phenylalanine - An unnatural derivative of L-phenylalanine that can be incorporated by recombinant methods into proteins allowing site-specific modification, e.g. by ‘click chemistry’, Staudinger reaction, or by UV-irradiation at 254 nm.  Compatible with downstream LC-MS⁄MS and MALDI MS analysis. Pair with DBCO or Alkyne

L-Azidohomoalanine - (AHA) is an amino acid analogue of methionine that contains a very small modification, specifically an azido moiety that can be fed to cultured cells and incorporated into proteins during active protein synthesis. Can be reacted in a CUAAC experiment with Alkyne, or DBCO in a copper free reaction.

L-Homopropargylglycine -  An amino acid analogue of methionine that contains a very small modification, specifically an alkyne moiety that can be fed to cultured cells and incorporated into proteins during active protein synthesis. Compatible with downstream LC-MS⁄MS and MALDI MS analysis. Detect with an azide reagent.