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Griffin:Antibody Basics: Difference between revisions
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A unique non-mammal source for research antibodies, polyclonal IgY from yolk sack is considered more humane than drawing blood. However success has been limited. | A unique non-mammal source for research antibodies, polyclonal IgY from yolk sack is considered more humane than drawing blood. However success has been limited. | ||
==Monoclonal Antibody purification== | |||
== | ===Hybridoma cell cutlure supernatant=== | ||
Centrifuge cultured medium at 1000 rpm for 5 minutes, collect supernatant into sterile containers, if necessary add sodium azide up to 0.2%. for several month storage, keep supernatant at 4C, otherwise, -20C. | |||
===Ascites fluid=== | |||
Hybridoma cell lines grown as ascites tumors in mice will frequently yield milligram quantities of monoclonal antibody per milliliter of ascites fluid. | |||
===Protein G purification=== | ===Protein G purification=== | ||
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7. Re-equilibrate the column in PBS with sodium azide. | 7. Re-equilibrate the column in PBS with sodium azide. | ||
==Polyclonal purification== | |||
===Peptide antigen affinity column=== | |||
===Protein A or G affinity column=== | |||
===Polyethylene Glycol Precipitation=== | ===Polyethylene Glycol Precipitation=== | ||
