Ascites purification

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Roshan's picture
Ascites purification

I want to purify a mouse IgG3 from crude ascites using protein A column. What is the ratio of the antibody to the gel? What is the best protocol to recover the antibody? How to wash and equilibrate the column before use?

What would have been different if I had IgG1 ascites instead of IgG3?

Please, if possible, respond quikly because I need to run my samples asap.

Guo-qiang Huang
Guo-qiang Huang's picture
Protein A is not the best

Protein A is not the best candidate for Mouse IgG3 purification in ascites. If you really want to choose affinity purification, Protien G might be better than protein A. Same suggestion for IgG1 ascites.

If I were you , I would like to choose Gel filtration or iron exchange to purify this antibody. the critical point is to avoid the cold precipitation of the IgG3.

Add colour 2 ur life
Add colour 2 ur life's picture
hi there plz go thru this