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  • Sample preparation

    CONCENTRATION

    Peptides: nM (p.e. Peptide 2KDa: ng/µL)

    Proteins: µM (p.e. protein 40 KDa: 2 µg/µL)

    INTERFERENCES

    Some solvents, buffers and detergents could cause noticeable degradation of the spectrum

    NO interference

    Tolerable (<50mM)

    Avoid

    TFA

    Formic acid

    DTT

    organic solvents,

    HCl

    acetic acid

    ammonium hydroxide

    HEPES
    MOPS

    TRIS

    Ammonium bicarbonate

    >1%Glycerol

    >1mM Sodium Azide

    >0.01 %SDS

    2 M Urea

    >2 M Guanidine

    >50 mM Phosphate buffer

    >0.1 %Detergents

    DMSO

    For protein identification (list all proteins contained in your gel or liquid samples), samples submitted for mass spectrometry analysis can come in:

    1) Liquid solutions

    Reduce contamination from keratin (or other proteins): In general the best results are always obtained from extremely clean samples. Contamination, primarily in the form of proteins present on the fingers and hair of investigators as well as dust in the lab should be avoided.

    Always wear gloves when handling samples

    2) Gel slices

    Reduce contamination from keratin (or other proteins): In general the best results are always obtained from extremely clean samples. Contamination, primarily in the form of proteins present on the fingers and hair of investigators as well as dust in the lab should be avoided. Some general recommendations:

    · Always wear gloves when handling samples

    · Never touch gels directly with bare fingers or leave gels exposed to dust in the lab atmosphere. This includes any apparatus involved in the preparation of gels or any surfaces that come into contact with gels.

    When cutting gel spots or slices always use a clean blade, the cost of a blade is far less than the cost of another protein ID analysis. Place the excised sample into a keratin-free, clean micro centrifuge tube. Tubes, pipette tips and other supplies that come into contact with the sample should come from closed plastic bags/boxes prior to use. We recommend to use glass containers and not to use it for other purposes that may contaminate the gel (for example Western-blot, since the BSA or milk protein could impregnate the gel).