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Advanced Techniques in Protein Analysis: From SDS-PAGE to Mass Spectrometry

This resource outlines various sophisticated methods for protein analysis, emphasizing their applications in biochemistry and molecular biology. Key topics include the use of electrophoretic mobility and frictional coefficients in polyacrylamide gel electrophoresis (PAGE), the principles of isoelectric focusing, and Mass Spectrometry techniques such as MALDI-TOF and ESI. Additionally, it covers methods like Edman degradation, cDNA cloning, and hybridoma cell ELISA. The insights provided improve sensitivity and resolution in protein studies and are essential for various research applications.

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Advanced Techniques in Protein Analysis: From SDS-PAGE to Mass Spectrometry

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  1. Excellent ed maps can be obtained by MAD phasing.

  2. Electrophoretic mobility Frictional coefficient Polyacrylamide gel Use fresh ammonium persulfate.

  3. Discontinuous buffer system SDS-PAGE Stacking gel Resolving gel

  4. log Mr SDS-PAGE 1. pI (isoelectric focusing) 2 Mr (SDS-PAGE)

  5. MWCO of dialysis tubing

  6. DEAE- CM- affinity column Gel filtration column: long Ion exchange column: medium Affinity column: small Concanavalin A

  7. Centrifugal force Sedimentation velocity Sedimentation coefficient 1 S=10-13 sec

  8. Mass under nondenaturing conditions

  9. MALDI TOF Matrix Assisted Laser Desorption Ionization Time-of-Flight ESI Mass Spectrum

  10. One cycle of Edman degradation Mixture of PTH-aa’s

  11. Performic acid oxidation and cleavage of -S-S- N-terminal sequencing Degenerate oligomer primer cDNA cloning DNA sequencing

  12. 4 Ca2+ binding motifs in calmodulin Hybridoma cell

  13. ELISA

  14. Imaging plate (IP) is 50-times more sensitive than X-ray film. Western blotting Actin filaments

  15. Gold cluster-conjugated antibody Secondary structure content

  16. Low resolution High resolution

  17. Spin=1/2

  18. Chemical shift When R < 5 A R

  19. X-ray vs NMR

  20. Natural Synthetic 항이뇨성의 DCC

  21. Merrifield solid-phase peptide synthesis

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