AHARIBO RNA / Protein Kit

AHARIBO RNA Kit

AHARIBO for RNA analysis is dedicated to the analysis of the translatome.

The product is based on an IMMAGINA technology called Minimally Invasive Non-canonical Amino acid

Tagging and Isolation of Ribosomes (Ribo-MINATI).

Ribo-MINATI technology is identification of whole de novo synthesized proteome and translatome following

the isolation of active ribosome.

 (Principle and Procedures)

Classical methods for translatome analysis such as polysome profiling or affinity purification-based

techniques followed by RNA-seq are characterized labor-intensive protocols and have so far led to relatively

poor correlations between mRNA and protein levels. AHARIBO for RNA analysis relies on a fast, low-input

protocol and can capture the dynamics of translation across different biological conditions. If combinedwith

AHARIBO for de novo proteome analysis, the kit allows for the parallel isolation and downstream analysis of

translated RNAs (by RNA-seq or qPCR) and the associated newly synthesized proteins.

AHARIBO for RNA analysis is based on the incorporation of non-canonical amino acid L-azidohomoalanine

(AHA) within ribosome-bound nascent peptide chains, followed by their stabilization on the ribosomes by

means of a blocking molecule and the final click chemistry- mediated pulldown of translational complexes

through the AHA-tagged nascent polypeptides.

Our core findings show that AHARIBO can enrich for (i) protein-coding RNAs, (ii) ribosome-associated

regulatory RNAs, and (iii) translated RNAs annotated as non-coding. AHARIBO-identified translatome

correlates better than total RNA with the respective protein levels in the dynamic framework of neuronal

differentiation of mouse embryonic stem cells. AHARIBO therefore represents an effective tool to

explorequantitative relationships between transcript and protein levels, offering a reliable and accurate

approach for taking snapshots of active translation processes.

Applications:

• Isolation of active polyribosomes and associated RNAs

• Easy and quick pull-down approach

• Parallel RNA and protein analysis if combines to AHARIBO for proteome

• Suitable for downstream RNAseq, qPCR and Ribo-seq.

  Protocol (PDF)

AHARIBO Protein Kit

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).

Ribo-MINATI technology is identification of whole de novo synthesized proteome and translatome following

the isolation of active ribosome.

 (Principle and Procedures)

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 proteinsynthesis

processes.

Applications:

• Isolation of nascent peptides associated with active polyribosomes

• Easy and quick pull-down approach

• Parallel RNA and protein analysis if combines to AHARIBO for RNA

• Suitable for downstream mass spec. and Western blot analysis.

  Protocol (PDF)

For the both items

• Shipping: dry ice and +4°C

• Storage Conditions: store components as indicated on the data sheet

• Shelf Life: 12 months