[Histonet] comments on Murine NK1.1(P136)

[Gayle Callis]

I was intrigued with the inquiry for this antibody and successful 
immunostaining.  A quick literature search turned up one publication, which 
must be the same one Sonya was reading(?).

Andrews DM, et el.  NK1.1+ cells and murine cytomegalovirus 
infections:  What happens in situ?, J Immunology 166:1796-1802, 2001.

This publication provided some excellent insight on what fixation worked 
best for this antibody.  They found the mouse had to be in vivo perfused 
with periodate lysine paraformaldehyde (PLP, McLean and Nakane, 1974, J 
Histochem Cytochem)  for the best results.  Immersion fixation was a poor 
choice of ex vivo frozen section from fresh tissue, snap frozen 
tissue.  They had a fixation chart and also a chart with results of 
fixation with staining on liver, spleen lung.   Also, it seemed to be 
important what strains of mice were used for this study.  They also 
quenched autofluorescence.  They did not indicate they did sucrose 
cryoprotection, but that would certainly make cryotomy easier with these 
tissues.

They did immunofluorescence staining with confocal laser scanning 
microscopy and had exceptional photographs of staining.

The authors commented that in situ identification of NK cells with this 
particular antibody clone (NK1.1, P 136) had been elusive "to date" (of 
this publication).   That is probably one reason BD Bioscience states in 
their technical data sheet is that this antibody does not work for 
immunohistochemical staining.

This antibody seems to one of the "picky" ones for tissue section work.

Good luck on getting this one to work.

Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610