Only cryo-preserve rapidly growing and dividing hybridomas (log phase hybridomas) that show more than 90% viability. Under sterile conditions, harvest the log-phase hybridoma cells in a centrifuge tube and perform a cell count using a hemacytometer and trypan blue. Centrifuge the cells at 200 g (or 1,000 rpm) for five minutes at room temperature. Carefully remove the medium and re-suspend the cell pellet in freezing medium (DMEM with 20% fetal bovine serum and 10% DMSO) to final cell density of 5-10x10^6 cells per ml and aliquot into cryovials at 1.0 ml/vial. Place the cryovials in cryo freezing containers or other suitable containers and place in freezer overnight at -80°C. The next day, transfer the cryo-vials to a liquid nitrogen tank. Alternatively, they can be shipped overnight if packed in a sufficient amount dry ice. Wear gloves and a facemask when working with liquid nitrogen tanks or frozen cryo-vials. If possible, send duplicate samples of each cell line.
Articles in this section
- I want to develop two phospho-specific pAbs, each one of them against one phospho-site. However, these two phosphorylated sites locate very close. Is it possible to get two specific pAbs that only recognize one phospho-site with no reaction to the other?
- Will Genscript ship some samples for test before final delivery?
- Why there is no affinity purification service for mouse pAb package?
- Why there is an extra cys in the antigen sequence you designed for me? Is it necessary to conjugate the peptide to a carrier protein and why?
- Why there is no immune-affinity purification option in mAb purification?
- Why is the antibody yield so much higher for the protein A-purified package verses the other purification methods?
- Why do you need to do KLH conjugation for my antigen? What carrier protein should I choose for conjugation?
- Why do I order test bleed?
- Why am I seeing multiple or unexpected bands on a Western with my affinity purified antibody?
- Which end of the peptide “C” should be added to for KLH conjugation?