• Cyberian's Gold

    Discussion Question:

    Which method is most suitable to identify an unknown protein on the basis of its molecular mass?

    Important Instructions:

    1. Your discussion must be based on logical facts.

    2. Your comments on the topic should not exceed 200-250 words.

    3. Support your answer with 5 well-versed statements.

    4. The GDB will remain open for 2 working days/ 48hours.

    5. Do not copy or exchange your answer with other students. Two identical / copied comments will be marked Zero (0) and may damage your grade in the course.

    6. Obnoxious or ignoble answer should be strictly avoided.

    7. Questions / queries related to the content of the GDB, which may be posted by the students on MDB or via e-mail, will not be replied till the due date of GDB is over.

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  • Cyberian's Gold

    Methods for Protein Analysis

    1. Protein Separation Methods
    2. Western Blotting
    3. Protein Identification Methods
      3A. Edman degradation
      3B. Mass spectrometry

    However, and perhaps remarkably, if as shown above we digest samples of ourprotein with as few as two or three different endoproteases with different
    specificities, we can usually use the resulting digestion patterns (again, analyzedby mass spectrometry to provide highly accurate determination of the fragmentmolecular weights) to produce a unique identification of our unknown protein:Again, mass spectrometry is uniquely well-suited for such analyses because itcan yield very accurate determinations of molecular weights from very even verysmall amounts of fragments resulting from the digestion of a particular protein.Techniques have now been developed by which proteins separated in two-dimensional gels can be digested within the gels and then injected directly into amass spectrometer for analysis of the resulting fragments.

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