Resources

Transgenic mouse lines

 

Mouse line Availability Reference genotyping primers
NG2DsRedBAC

 

Jackson Laboratory

Stock 008241

 

Zhu et al., Development 2008

DsRedF: 5′-ccgaggacgtcatcaagg

 

DsRedR: 5′-gagccgtactggaactgg

 

Product: 196 bp

NG2CreBAC

(cryopreserved)

Jackson Laboratory

stock 008533

Zhu et al., Development 2008

Note: breed through females only

CreF37: 5′-ccgtacaccaaaatttgcc

 

CreR238: 5′-atcgcgaacatcttcagg

 

Product: 202 bp

NG2creERTMBAC Jackson Laboratory

stock 008538

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.

Methods

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

Anti-NG2 antibodies

  1. EMD Millipore AB5320 rabbit anti-NG2 chondroitin sulfate proteoglycan antibody, 1:500 dilution on cultures and fixed mouse and rat brain tissue

 

Anti-Pdgfra antibodies

  1. R&D Systems, AF1062, goat anti-mouse Pdgfra antibody 1:2000 dilution, good for fixed mouse brain tissue
  2. BD Sciences, 558774, CD140a rat monoclonal anti-mouse Pdgfra antibody, IgG2a