Which markers best differentiate bovine neutrophils, macrophages and lymphocytes for flow cytometry cell sorting?

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Ina Laura Pieper

Hi Serhat! I have a similar problem, I want to study these in bovine blood with flow cytometry. I have found many CD14-antibodies that cross-react with bovine monocytes: 61D3, HCD14, Tuk4, and M5E2. These are supplied by many vendors, so you shouldn't have a problem finding the right fluorchrome for your panel. For B-cells, you could use CC21 (BoCD21). For T-cells you can use CC8 (BoCD4) and CC63 (BoCD8). They're from Sopp P, Vet Immunol Immunopathol, 2001, but I don't know where you buy them. My main issue is the neutrophils - I really need a lineage marker for these. If you find one, please let me know :)

Best regards,

Ina Laura

Hi again, Mab IL-A 110 seems to target bovine neutrophils. I haven't found a supplier. If you find out how to get it, please let me know.

Thanks,

Serhat Sevli

Hi Ina,

I havent heard the IL-A110 before, what is the CD name for that? I have chosen CH138A, CD11b, CD11a/CD18, and CD62L. I plan to separate and analyse these populations and determine which is best.

You may ask William Davis at Veterinary Medicine Department of Washington State University. He helped me very much while choosing those four markers. http://www.vetmed.wsu.edu/depts-MoAb/index.aspx

Thanks for your answer by the way.

Hi Serhat! I contacted William yesterday too, he recommended CH138A. Have you tried it yet? I was planning to purchase it and see how it works, but if you've already tried it I'd love to hear what you think. Does it increase/decrease in expression if the neutrophils are activated? The IL-A 110 is referenced in this article (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC440617/) and was a gift from Jan Naessens. He's on Researchgate so I'll ask him if I can try the antibody :)

Best regards, Ina Laura

Xichen Zhang

Hi Serhat and Ina

Here is a very late follower. Just wondering have you guys have any concluded information about the antibody or the supplier for the phenotyping of bovine lymphocytes and neutrophils.

Looking forward. Many thanks.

Xichen

Hello Xichen Zhang

Yes we have some results about neutrophil markers but unable to tell currently because of patent application.

Macrophage marker is obviously CD14. I also could not find another.

Thanks and best wishes

Serhat

Abhi Jayasimhan

Hi Serhat,

I am interested to know if your CH138A worked good for neutrophils? Does CH138A binds to all granulocytes or is it specific for only Neutrophils?

Mohammad Asad

Hi,

Below paper may be of your interest and helpful.

Article Phenotype and Function of CD209+ Bovine Blood Dendritic Cell...

Regards,

Asad