Resources

Transgenic mouse lines

NG2DsRedBAC

Jackson Laboratory, Stock 008241

Zhu et al., Development 2008

Genotyping primers

DsRedF: 5′-ccgaggacgtcatcaagg

DsRedR: 5′-gagccgtactggaactgg

Product: 196 bp

 

NG2CreBAC

Jackson Laboratory, stock 008533   (cryopreserved)

Zhu et al., Development 2008

Genotyping primers

CreF37: 5′-ccgtacaccaaaatttgcc

CreR238: 5′-atcgcgaacatcttcagg

Product: 202 bp

 

NG2creERTMBAC

Jackson Laboratory, stock 008538

Zhu et al., Development 2011

Genotyping primers

same as for NG2CreBAC

 

Optimized fixation for NG2 and PDGFRα immunofluorescence

To preserve NG2 and PDGFRα 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.

See Pfeiffer and Nishiyama, Cells 2021

Methods

 

Sources of antibodies for detecting NG2 and PDGFRα 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-DGFRα antibodies

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