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文献和实验Notes on Making Rat x Y3 Monoclonal Antibody Producing Hybridomas
Although Y3/Ag1.2.3 has its own myeloma light chain, their are numerous characteristics that make this a better parent line for making hybridomas than the 'Y0' chain loss variants or using NS0 to make rat x mouse hybrids. These are: a) Hybridomas
To fix, 1/10 volume of 37 formaldehyde is added to the culture, which is shaken for a further 40 minutes at room temperature. Cells are then recovered by centrifugation at 1400 X g for 2 minutes, and washed (i.e. gently resuspended then recovered
Anti-GFP/BrdU In Situ Immunostaining
by adding 1 mL PBS, 3' each. Add 0.3 mL rabbit anti-GFP (1:200 v/v, plus 1% goat serum in PBS) and incubate 1 hr. at room temp. Wash 3x in PBS. Post-fix in 1 mL 4% paraformaldehyde x15', room temp. Wash 3x in PBS. Denature DNA with 1 mL
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