AHARIBO for de novo proteome analysis is dedicated to the analysis of newly synthesized proteins. The product is based on an IMMAGINA technology called Minimally Invasive Non-canonical Amino acid Tagging and Isolation of Ribosomes (Ribo-MINATI).
Classical methods for de novo proteome analysis such as pSILAC or affinity purification-based techniques (e.g. PUNCH-P) followed by LC-MS are characterized by labor-intensive protocols with relatively poor correlations between mRNA and protein levels. AHARIBO for de novo proteome analysis relies on a fast, low-input protocol and can capture the dynamics of protein synthesis across different biological conditions. If combined with AHARIBO for RNA analysis, the kit allows for the parallel isolation and downstream analysis of translated RNAs and the associated newly synthesized proteins (e.g. by mass spectrometry).
AHARIBO for de novo proteome analysis is based on the incorporation of non-canonical amino acid L-azidohomoalanine (AHA) within ribosome-bound nascent peptides, followed by their stabilization on the ribosomes by means of a blocking molecule and the final click chemistry-mediated pulldown of the AHA-tagged nascent polypeptides.
Our core findings show that AHARIBO-identified proteome correlates better than total proteome with the respective mRNA levels in the dynamic framework of neuronal differentiation of mouse embryonic stem cells. AHARIBO therefore represents an effective tool to explore quantitative relationships between transcript and protein levels, offering a reliable and accurate approach for taking snapshots of ongoing protein synthesis processes.
Shipping: dry ice and +4°C
Storage Conditions: store components as indicated on the data sheet
Shelf Life: 12 months
The kit contains reagents for 6 experiments
Reference. L. Minati et al. One-shot analysis of translated mammalian lncRNAs with AHARIBO bioRxiv, April 20, 2020
Protocol AHARIBO protein