Download Fluorescence Imaging

FLUORESCENCE IMAGING
Quantification .................................................................................................33
One-dimensional gel/blot analysis.............................................................33
Array and microplate analysis ..................................................................35
Two-dimensional protein gel analysis .......................................................36
Background correction ....................................................................................36
Image processing tools.....................................................................................40
Amersham Pharmacia Biotech image analysis software ...................................41
Chapter 5: Fluorescence applications using
Amersham Pharmacia Biotech imaging systems ...........................................45
Introduction.....................................................................................................45
Detection of nucleic acids in gel ......................................................................45
Nucleic acid gel stains...............................................................................45
Instrument compatibility...........................................................................47
Typical protocol........................................................................................47
Expected results ........................................................................................49
Detection of proteins in gel..............................................................................51
Protein gel stains.......................................................................................51
Instrument compatibility...........................................................................52
Typical protocols ......................................................................................53
Protein detection in one-dimensional gels ..........................................53
Protein detection in two-dimensional gels ..........................................55
Expected results ........................................................................................58
Quantification of nucleic acids in solution ......................................................59
Dyes for quantification of nucleic acids in solution .................................59
Instrument compatibility...........................................................................60
Typical protocol........................................................................................60
Expected results ........................................................................................62
Quantification of proteins in solution..............................................................63
Dyes for quantification of proteins in solution .........................................63
Instrument compatibility...........................................................................64
Typical protocol........................................................................................64
Expected results ........................................................................................66
Southern and Northern blotting ......................................................................67
Fluorogenic substrates for Southern and Northern detection....................67
Instrument compatibility...........................................................................69
Typical protocol........................................................................................69
Expected results ........................................................................................72
Western blotting ..............................................................................................73
Western detection strategies ......................................................................73
Enzyme-amplified detection (chemifluorescence)................................73
Direct fluorescent detection................................................................75
Total protein stains for Western blots ................................................75
Instrument compatibility...........................................................................76
Typical protocols ......................................................................................77
Western blotting using a fluorogenic substrate...................................77
Western blotting using a fluorochrome-conjugated antibody .............80
Expected results ........................................................................................82
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