mAb characterization

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mAb characterization

Postby gene26 » Aug 23 2017 4:49 pm

After cell fusion and screening for hybrids that secrete mAb against the antigen. Usually, plates are coated with antigen concentration 1ug/well and 100ul of cell culture supernatant as a sample. Is it necessary to dilute supernatant?

For sensitivity: Sandwich ELISA is employed with mAbs from two different clones that secrete against the same antigen. mAb from one clone will be used as capture Ab and from the second clone as primary Ab.

Now, what concentration of purified mAB should I use for capture Ab 1000ng or 500ng or 250ng or try them all? What is the correct way to pick appropriate concentration purely based on observation?

and my antigen is a protein (both modified and native) the sample concentration should vary from for 1000ng -10ng?

lastly primary antibody concentration range?
gene26
Prolific Post-Master
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