Preparing samples for N- and C-terminal sequencing by MALDI ISD

Alphalyse
Feb 22. 2014

Question:

How do I prepare samples for MALDI ISD analysis?

I am interested in your N and C-terminal sequencing using MALDI ISD. How do I prepare the sample for analysis?

Answer:

MALDI In-Source Decay (ISD) is used for confirmation of the N- and C-terminal of known proteins, therefore you would need to supply us with the sequence of the protein you are working with so we can confirm the sequence of the two termini [1-3].

We require the samples to be in liquid form with a purity >80%. We need 20 ug of material to do the analysis. If you can provide more material, it would be ideal. The more material we receive the better the quality of the results will be.

When you submit your samples you should list what is in the buffer you are using. Please let us know if there is glycerol in the sample as we need to remove this before placing the sample in the mass spec. Purify the sample by reversed-phase HPLC.

You should note that we perform the N- and C-terminal sequencing by MALDI ISD simultaneously on each sample.

Read more about the Alphalyse N- and C-terminal sequencing service >>>

References:

[1]          Hillenkamp et al:Matrix-assisted laser desorption/ionization mass spectrometry of biopolymers“, Analytical Chemistry, 1991

[2]          Debois et al: “MALDI-In Source Decay Applied to Mass Spectrometry Imaging: A New Tool for Protein Identification“, Analytical Chemistry2010

[3]          Demeure et al: New Advances in the Understanding of the In-Source Decay Fragmentation of Peptides in MALDI-TOF-MS“, Journal of the American Society for Mass Spectrometry2010

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About Alphalyse

Alphalyse uses expertise in the field of protein chemistry and bioinformatics, combined with top-of-the-line mass spectrometry equipment, to provide protein analysis services globally.

Alphalyse aims to deliver high quality data and customer service to all of our clients in a fast and convenient way.

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