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Notes on Making Rat x Y3 Monoclonal Antibody Producing Hybridomas

of one rat spleen (approx. 10**8 cells) together with 6 x 10**7 Y3/Ag1.2.3 per fusion (the other half of the spleen cells can be frozen as a backup in 90% FCS +10% DMS0). c) Try and keep all cells, PEG and medium components at 37 deg. C

ANTIBODY BINDING TO PROTEIN A AND PROTEIN G

2a ++++ ++++ Mouse IgG2b +++ +++ Mouse IgG3 ++ +++ Rat IgG1 - + Rat IgG2a - ++++ Rat IgG2b - ++ Rat IgG2c

secondary antibody review -- data from 99 publications

western blot   test Benzonase activity in PMCA reactions containing biotin-labeled oligo(dT),   17 goat IgG   FITC flow cytometry 1:250   Jackson ImmunoResearch Laboratories 17 goat     AP ELISA 1:1000     18       HRP western blot     Cell