Transgenic mouse lines
|Mouse line||Availability||Reference||genotyping primers|
|Jackson Laboratory||DsRedF: 5′-ccgaggacgtcatcaagg
Product: 196 bp
|Jackson Laboratory||Zhu et al., Development 2008
Note: breed through females only
Product: 202 bp
|NG2creERTMBAC||Jackson Laboratory||Zhu et al., Development 2011||Same as for NG2creBAC|
Optimized fixation for NG2 and Pdgfra immunofluorescence
To preserve NG2 and Pdgfra antigen for immunofluorescence, it is important not to overfix the tissue.
Perfuse animals with 4% paraformaldehyde in PLP (periodate-lysine-paraformaldehyde) fixative at room temperature. For surface antigents, 3% paraformaldehyde works better. With 2% antigen is preserved better but tissue is soft and difficult to handle, especially CNS tissue from young animals.
See Mori et al., 2009 for an illustration of the effects of PFA concentration on NG2 immunofluorescence signal
Sources of antibodies for detecting NG2 and Pdgfra on mouse tissue
- EMD Millipore AB5320 rabbit anti-NG2 chondroitin sulfate proteoglycan antibody, 1:500 dilution on cultures and fixed mouse and rat brain tissue
- R&D Systems, AF1062, goat anti-mouse Pdgfra antibody 1:2000 dilution, good for fixed mouse brain tissue
- BD Sciences, 558774, CD140a rat monoclonal anti-mouse Pdgfra antibody, IgG2a