Q1: No positive clones during hybridoma fusion screening.
The number of plates made after fusion can be increased to between 10 and 20. At the same time, the concentration of HAT medium can be appropriately increased, and other batches of Sp2/0 cells can be used.
Q2: There is precipitate in the product. Can it be used?
Shake well and use it normally. After shaking well, there is still some sediment at the bottom, but it does not affect its use. It can be dissolved in a 37-degree water bath appropriately, and most of it can be redissolved.
Q3: What is the best way to dilute 50x HAT selection medium to 1x?
The proportion of each component in the HAT medium for preparation: 15% fetal bovine serum, 10% added factor, 1×HAT selective medium, 1× double antibiotic Penicillin-Streptomycin, dissolved in incomplete 1640 medium. HAT medium cannot replace serum, so if diluted with sterile water, it cannot provide the nutrients and growth factors required for cell growth.
Q4: Is it normal to have yellowish-green/white precipitate?
This is supersaturated precipitation, which is normal. If the precipitated substance contains A, it will be yellowish-green and does not affect its use. It can be dissolved in a 37℃ water bath
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