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文献和实验(determine experimentally). A dilution series of 1:100 to 1:10,000 should cover it. Incubate at RT for 1 hr or longer, depending on the antibody. Sometimes an overnight incubation is helpful. Jon''s anti-actin Guinea pig antibodies from animal#1 are best
treat the cells with 30 ul 10 mg/ml Zymolyase (100T) at 30¡C for anywhere between 10-30 min or more. Examine cells on a phase microscope. When cells are dark and mishapen, you have gone way too far, if bright and refractile they probably n eed
Yeast IF without Dehydration--酵母免疫荧光（无甲醇脱水方法)
) and once in the same buffer containing 1.2M sorbitol. Be very gentle with the cells! Resuspend in 0.5 mls sorbitol buffer. The cells can be stored at this point overnight at 4¡C. Zymolyase treat the cells with 30 ul 10 mg/ml Zymolyase (100T) at 30¡C
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