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文献和实验RUVKUN Antibody Staining Protocol revised 10/29/90
RUVKUN Antibody Staining Protocol revised 10/29/90 Fixation: 1. C. elegans N2 grown in liquid or plates. Harvest worms. Wash for 2 hours at room temp. in 1X M9 or PBS. 2. Fix in 5 ml of FRESH THAT DAY 1% paraformaldehyde:
the antibody, load the affigel concentrate onto the column and hold at room temperature for 10 minutes.(15) Wash with 50 ml 10 mM Tris 7.5 and then wash with 50 ml 10 mM Tris 7.5/500 mM NaCl. Elute with 5 ml 100 mM Glycine pH 2.5 into 1 ml 1M Tris 8.0.(16
. The first is to Dialyze the coupled antibody against PBS for 24 hours with 3 changes of PBS. Doing this will not change concentration of original antibody (recommended for antibody with a concentration less than 1mg/mL). The second way is to use a PD-10 column. Using
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