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Preparation of Samples Prior To Mass Spectrometry

Preparation of Samples Prior To Mass Spectrometry - Mass Spectrometry Forum

Preparation of Samples Prior To Mass Spectrometry - Discuss and post questions about LC-MS, Q-STAR, MALDI-TOF, and other Mass Spectrometry Questions in the Mass Spec Forum.


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  #1  
Old 06-15-2006, 09:12 PM
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Arrow Preparation of Samples Prior To Mass Spectrometry



Hey all,

I am trying to prepare samples for Mass Spectrometry. I know that desalting and protein concentration are important, but what else is important?

What buffers are compatible with mass spec?

thank U
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Old 06-15-2006, 09:16 PM
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Default Re: Preparation of Samples Prior To Mass Spectrometry

Also are there any straight and fast rules?
here are some that i know:

Avoid Buffers such as :
TRIS, CHAPS, PBS, and HEPES

Avoid Salts:
e.g. Na sodium, K potassium, P phosphate, citrate, perchlorate

Avoid surfactants and detergents

What is acceptable then?
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Old 06-29-2006, 04:36 PM
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Default Re: Preparation of Samples Prior To Mass Spectrometry

Looks good so far. I always dialyze in Ammonium Bicarbonate. very compatible!
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Old 08-17-2006, 09:32 AM
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Default Re: Preparation of Samples Prior To Mass Spectrometry

For me.. i would jst freeze dry the sample then re-dissolve it in acetic acid.. if not then in 0.04% aqueous TFA.. If not you may try using a centrifugal membrane to desalt your sample and concentrate it down..
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Old 08-17-2006, 11:23 AM
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Default Re: Preparation of Samples Prior To Mass Spectrometry

Hi
i am giving below a link of virtual mass spectrometry laboratory. and here they have given the procedure for sample preparation in the form of video, i hope you will find it very informative for you.

[Only registered users see links. ]

best regards
aftab ahmad
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Old 08-17-2006, 11:57 PM
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Default Re: Preparation of Samples Prior To Mass Spectrometry

Wow! great find Aftab!
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Old 09-19-2006, 02:39 AM
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Thumbs down Help me please???

Hello every body.

I am a new member. My name is Minh. I come from proteomics Lab, Institute of Biotechnology, Hanoi, Viet nam.

I have some difficulties in a proteomics application to identify HBs Ag (from a recombinant vaccine sample). The sample comprises: HBs Ag 20ug/ml; Al(OH)3 < 1.25mg/ml; Themerosal< 0.1<%; formaldehyde<0.01%.
I tried several methods:
1. SDS-PAGE of 100ul sample (dryness), 1 observed band (Mw appro. 22kda) then was cut and followed by ingel-digestion and ESI-1DLCMS/MS identification (AB_QSTAR XL system).
2. Direct trypsin digestion (insolution digestion) of 100ul sample (dryness), following by ESI-1DLCMS/MS identification.
I spent more than 2 weeks on doing this work. now, I still do not identify the protein of interest.
I think that Al(OH)3 may impact on ability of trypsin digestion. But I don't know how to remove the impact.

Please suggest me how to solve this problem.
I am looking forward to hearing from you soon.

Thanks and best regards.

Pham Dinh Minh
Protein Biochemistry Lab, Institute of Biotechnology
Vietnamese Academy of Science & Technology

18 Hoang Quoc Viet Street, Cau Giay Dist., Ha Noi , Viet Nam

Tel: (+84)-4-7561903 Fax: (+84)-4-8363144 Emai: [Only registered users see links. ]
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Old 10-19-2006, 10:49 PM
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Default Re: Preparation of Samples Prior To Mass Spectrometry

I would try dialyzing your sample Minphd in trypsin compatible buffer and trying digestion again and reanalysis.

Maybe your buffer is affecting the trypsin and/or the mass spectrometer machine.
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Old 01-29-2007, 10:43 PM
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Default Re: Preparation of Samples Prior To Mass Spectrometry

a suggestion to you.. You may consider trying to use the RapiGest solution from Waters.. it helps with the digestion..
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