Polyclonal antibodies against the extracellular (Millipore Cat# AB1554 RRID:AB_90760) and cytoplasmic (Promega Cat# G3231 RRID:AB_430853) domains of the low-affinity neurotrophin receptor p75NTR (both at 1:400 dilution) were used to identify Schwann cells in vitro (Provenzano et al., 2011 (link); Whitlon et al., 2009 (link)), determine their specific location within synapse cultures, and their alignment to neighboring spiral ganglion neurons. Western blot and immunocytochemical analysis of antibody specificity showed the expected lack of protein in p75NTR−/− animals compared to wild type controls (Gehler et al., 2004 (link); Shulga et al., 2012 (link); Wagner et al., 2011 (link)).
Protocol detail
Identifying Schwann Cells in Synapse Cultures
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Animals
Antibodies
Antibody specificity
Cytoplasmic
Dilution
Neurons
P75ntr
Promega
Protein
Schwann cells
Spiral ganglion
Synapse
Western blot
Corresponding Organization : Rutgers, The State University of New Jersey
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Variable analysis
independent variables
- Genotype (p75^NTR−/− vs. wild type)
- Protein expression levels of p75^NTR
- Schwann cell location within synapse cultures
- Alignment of Schwann cells to neighboring spiral ganglion neurons
- Antibody dilution (1:400) for both extracellular and cytoplasmic domains of p75^NTR
- Positive control: Wild-type animals
- Negative control: p75^NTR−/− animals
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