In my hybridoma culture plate, fused cells are not forming colonies/clumps even after 24 days (Cells as single-image attached). After 12 days I shifted from HAT media to HT and is not showing much proliferation and clump formation. I followed the protocol from Antibody Methods and Protocols, Methods in Molecular Biology (Zhang, 2012).
Ibrahim Hatipoglu
Dear Mahesh,
I think your problem about fusion step. When you fused splenocytes and myeloma cells with PEG4000, you should be gentle.
good luck
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