Sci. STKE, 20 June 2000
A corrected PDF is available from http://stke.sciencemag.org/cgi/reprint/OC_sigtrans;2000/37/pl1. The original PDF for this file had the following errors:
1. Page 2, Fig.1
2. Page 3, Recipe 8
"10 mM" should have read "10 mM in 50mM NH
3. Page 4, Instruction 11
The original directions were modified to read:
"Add 200 µg of agarose-conjugated 4G10 monoclonal antibody and 75 µg of the antiphosphotyrosine antibody RC20 to the supernatant. Add 300 µl of settled-streptavidin-conjugated agarose beads to capture this biotinylated antibody. Incubate for 6 hours at 4°C. This step can also be performed overnight.
Note: Because different monoclonal antiphosphotyrosine antibodies immunoprecipitate slightly different sets of tyrosine phosphorylated proteins, we routinely add the second antiphosphotyrosine antibody (RC20) during the immunoprecipitation step.
If the starting amount of culture volume is more than 1 liter, we recommend that the immunoprecipitation be done in multiple batches instead of scaling up the amount of antibody added."
4. Page 7, Fig. 5 in the section called "Searches against EST database."
"Impossible" should read "Difficult"
5. Page 10, Fig. 6 in the section called "Problem amino acids"
"K and E can be distinguished..." should read "K and Q can be distinguished..."
6. Reference 27 should read
Mamyrin, B. A. (1994) Laser-assisted reflectron time-of-flight mass spectrometry. Int. J. Mass Spectrum Ion Processes 131: 1-19.
7. Reference 30 should read
Vorm, O., Roepstorff, P., and Mann, M. (1994) Improved resolution and very high sensitivity in MALDITOF of matrix surfaces made by fast evaporation. Anal. Chem. 66: 3281-3287.
Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882