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UC Davis/NIH NeuroMab Facility Protocols

NeuroMabs and other mouse mAbs are typically available in two forms:

TC Supernatant: conditioned bovine serum-containing media harvested from hybridoma cells cultured in vitro. Typically contains 20-40 ug/mL mouse monoclonal lgG plus 10 ug/mL bovine lgG and other bovine serum components. Can contain substantially higher amounts of mouse monoclonal lgG. Recommended working dilutions for TC supernatants are 1:2-1:20 for immunoperoxidase-based immunohistochemistry, and 1:2-1:10 for immunofluorescence-based immunohistochemistry and immunocytochemistry, and immunoblotting.

Purified lgG: Predominantly mouse monoclonal lgG but may contain trace amounts of other mouse lgG. Recommended working concentrations for purified lgGs are 0.1-1 ug/mL for immunoperoxidase-based immunohistochemistry and immunoblotting, and 1-10 ug/mL for immunofluorescence-based immunohistochemistry and immunocytochemistry.

These recommended working dilutions/concentrations are meant to be a starting point for optimization based on your specific application.  Further suggestions can be found within individual protocols. 

Optimize immunolabeling with NeuroMabs and other mouse monoclonal antibodies by employing appropriate subclass-specific secondary antibodies. See article here.

Immunofluorescence immunocytochemistry of cultured cells (last updated September 2016)

Preparation of mammalian brain fractions (last updated September 2016) 

Immunoblot labeling (last updated September 2016)

Perfusion fixation of rat brain (last updated October 2016)

Perfusion fixation of mouse brain (last updated October 2016)

Immunofluorescence labeling of brain sections (last updated October 2016)

Immunoperoxidase labeling of brain sections (last updated October 2016)

Antigen retrieval of rat brain sections (last updated September 2016)

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