Antibodies to ID mouse neutrophils and monocytes

[SCIPHILO]

Hi there,

I am looking for specific antibodies which can help me to differentiate between mouse neutrophils and monocytes in FACS or Luminex assays. So far, I have come across the following from eBioscience:

Neutrophil subset: anti-mouse CD177
Monocyte/myeloid: anti-mouse CDw329

Does anybody know a more specific product(s)? Or can suggest another manufacturer?

Regards,

SP

[DoctorStein]

If I am not mistaken, the equipment is able to distinguish between neutrophils and monocytes. The simpler equipment like Flow cytometer also able to distinguish between them quite easily. Or maybe I mistook your question here?

[SCIPHILO]

Thank you for your reply. I am a novice in this field; however I have been told that since neutrophils and monocytes are both highly granulated, they are difficult to be distinguished just by Flow Cytometry. Hence, sorting by Luminex which uses specific antibodies attached to microspheres could be a good tool to separate them.

[DoctorStein]

I have just used it few months ago to perform leucocyte differential numbers and I got no problem on distinguishing between neutrophil and monocyte (even you can do this manually just like what I did by blood smear preparation and light microscope when the equipment was broken here ) Those cells have different size and different granules, which the equipment can recognize it by "forward scatter" (FSC) and by "side scatter" (SSC), respectively. I do not use any antibodies, which is so damn expensive here, to perform this routine analysis.

Actually, FACS is kind of flow cytometer

[SCIPHILO]

Thank you DoctorStein.

[biohazard]

It all depends on how certain you want to be about the cell %'s - if you only need numbers, simply put proper gates in the FSC / SSC dot plot and it should usually be enough. However, if you want, for example, to be sure that your neutrophils don't contain any contaminating monocytes (or vice versa), you need to label them with antibodies. Some of the granular monocytes can sometimes appear in the neutrophil population and same can happen the other way round.

Now, what comes to specific markers for mouse cells, I cannot say for certain atm. But I'll post here if I come up with something.

[SCIPHILO]

Thank you for your input. I need to use antibodies for precisely the same reason that you mentioned.

[DoctorStein]

What about this thing:
- NIMP-R14
The monoclonal antibody NIMP-R14 has been succesfully used to stain neutrophils for fluorescent activated cell sorting and in frozen and paraffin sections.

- Sa14-2
Sa14-2 is a monoclonal antibody that reacts with the mouse monocyte marker CD14.