Summary 1

ORIGINAL QUESTION: Anyone have suggestions for canine and porcine antibodies and cytokines? (Posted 1/31/17) by J. Paul Robinson, Purdue)

Here are the responses I received and contact information for respondents.

From Ashley Hartley

Manuscript of interest:

Chemokine receptor 7 (CCR7)-expression and IFN production define vaccine-specific canine T-cell subsets Ashley N. Hartleya,b,1, Rick L. Tarletona, Veterinary Immunology and Immunopathology 164 (2015) 127–136 

c,∗ a Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA, 30602, USA b Department of Infectious Diseases, University of Georgia, Athens, GA, 30602, USA
c Department of Cellular Biology, University of Georgia, Athens, GA, 30602, USA

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From Dirk Werling:

https://www.bio-rad-antibodies.com/domestic-animals.html

Similarly, Washington State University offers a whole lot:

http://vmp.vetmed.wsu.edu/resources/monoclonal-antibody-center

Both have limited sets to canine cytokines as well, but if you want

recombinant canine cytokines, the widest offer is with Kingfisher

https://www.kingfisherbiotech.com/category/ProductType_PROTEIN/

Hope that helps?

If you want other molecules/CD markers, please get in touch, I might be able to point you in the right direction or help with protocol for

FACS/ELISA/ELISPot

Best wishes

Dirk Werling

Professor of Molecular Immunology

Royal Veterinary College

University of London

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FROM: Rodolfo D. Vicetti Miguel

We have had good experiences with antibodies from eBioscience and Biorad:

http://us.ebioscience.com/application/flow-cytometry/canine-antibodies.htm

https://www.bio-rad-antibodies.com/search.html?searchType=BASIC&searchTerm=anti%20Canine&cat=&Filter1Name=TargetSpecies&Filter1Value=Dog&filterCount=1&type=

We have used antibodies from eBioscience for T cell markers (CD4, CD8, CD5) and for B cells (CD79a, intracellular marker, clone HM47). Also, we have used anti-CD21 (surface, clone CA2.1D6) from Biorad for B cells. If you are activating canine B cells, I would strongly recommend to use both anti-CD21 and anti-CD79a, if possible. For IFN-g detection, we have used clone CC302 from Biorad.

Only issue we found was big spillover into PE with PE-Cy7 conjugates from eBioscience.

If you are doing ELISPOTs, you may want to look into these mAbs:

https://www.mabtech.com/products/3115-3-anti-bovine-ifn-%CE%B3-mab-bifn%CE%B3-i-purified

https://www.mabtech.com/products/3115-6-anti-bovine-ifn-%CE%B3-mab-pan-biotinylated

Finally, I am attaching a paper that I think would help you. We have not used all the antibodies they used, but we have been able to reproduce some of their results.

Please let me know if you have any questions.

Good luck with your project.

Best,

Rodolfo D. Vicetti Miguel, M.D.
Basic Life Science Research Scientist
Department of Comparative Medicine
Stanford University School of Medicine
300 Pasteur Drive, Edwards R312
Stanford CA  94305
650.721.8961

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FROM Jonathon Fogle

We perform a good bit of immunophenotyping for dogs in our clinical lab and in our research lab.  My email is here if you'd like, I'm happy to share the antibodies we use day to day for our studies.  

Jonathan

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FROM Katie Quarfoth <Katie.Quarfoth@bio-techne.com> wrote:

Hi Paul – We have not been in contact before, but I wanted to reach out and introduce myself as one of your sales rep for R&D Systems.  Myself, along with my colleague Jen Souvignier are your sales reps and we are happy to help with anything you may need for your research.  I wanted to let you know of our extensive antibody offering in case you weren’t aware.  R&D Systems along with our sister brand Novus Biologics offers a huge list of antibodies – many for porcine and canine. 

Here is our complete list of R&D Systems Porcine antibodies:

https://www.rndsystems.com/search?keywords=porcine&category=Primary%20Antibodies&species=Porcine

and those validated specifically for flow cytometry:

https://www.rndsystems.com/search?keywords=porcine&category=Primary%20Antibodies&species=Porcine&rnd_applications=Flow%20Cytometry

Here is our complete list of R&D Systems Canine antibodies:

https://www.rndsystems.com/search?keywords=canine&category=Primary%20Antibodies

and those validated specifically for flow cytometry:

https://www.rndsystems.com/search?keywords=canine&category=Primary%20Antibodies&rnd_applications=Flow%20Cytometry

Here is the complete list of Novus Porcine & Canine Antibodies:

https://www.novusbio.com/search?keywords=novus&category=Primary%20Antibodies&species=Canine|Porcine

and those validated specifically for flow cytometry:

https://www.novusbio.com/search?keywords=novus&category=Primary%20Antibodies&species=Canine|Porcine&applications=Flow%20Cytometry

Let us know if there is an antibody you are specifically looking for.  We are also happy to offer discounting for you to try out any of our antibodies.  Feel free to contact us anytime and we look forward to working with you!

FROM Davis Seelig

FROM Bill Davis

We have some mAbs to canine LDMs http://vmp.vetmed.wsu.edu/resources/monoclonal-antibody-center.  Kingfisher Biotech has an array of canine cytokines plus they are a distributor of our mAbs http://www.kingfisherbiotech.com/category/species_canine/.  I hope this helps.

Bill  

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FROM Kerry White

FROM Eric van Burren

We have also been contacted recently regarding canine phenotyping antibodies. The project is still in the planning phase.

 The traditional response is that there are very few commercial antibodies specific for canine. These are from AbD Serotec (now BioRad), Abcam, ebioscience, and others. Using these traditional antibodies the palette is rather limited and immunophenotyping panels look a lot like unsophisticated human/mouse panels from 25+ years ago. (An added frustration is that perhaps the best B cell marker is intracellular.)

A non-traditional approach is exemplified by Christine Peterson's group at Iowa (see below). They simply used human/mouse antibodies that cross react with canine. Presumably these have been properly validated but I haven't seen any evidence of this.

 Schaut RG, Lamb IM, Toepp AJ, Scott B, Mendes-Aguiar CO, Coutinho JF, Jeronimo SM, Wilson ME, Harty JT, Waldschmidt TJ, Petersen CA. Regulatory IgDhi B Cells Suppress T Cell Function via IL-10 and PD-L1 during Progressive Visceral Leishmaniasis. J Immunol. 2016 May 15;196(10):4100-9. doi: 10.4049/jimmunol.1502678. PubMed PMID: 27076677; PubMed Central PMCID: PMC4868652.

 Eric Van Buren, Flow Cytometry Services manager

Microscopy, Imaging and Cytometry Resources Core

Barbara Ann Karmanos Cancer Institute

Wayne State University

(313)576-8341
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FROM Dorothee Bienzle

There are a couple of veterinary academic labs that offer immunophenotyping of canine leukocytes with various subsets of the antibodies below. Not sure if you are looking for more detailed protocols? There are also several people who have used the Luminex/Millipore platform for assessing canine cytokines. 

Dorothee

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Dorothee Bienzle, DVM, PhD, Dipl. ACVP

Professor of Veterinary Pathology

Department of Pathobiology

University of Guelph

Room 3822, Building 89

419 Gordon St.

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FROM Sarah Schuett

For canine phenotyping antibodies we use ebioscience. They have a lot of great choices available. Also, Novus; their antibodies are pricey but we have great results. I'd steer clear of serotec (now biorad) if possible as we've had many issues of the antibodies not properly working. We've got a decent basic panel for T and B cells here at NC State Vet School as we've got a diagnostic lab, if you need any further info just contact me directly.

Thanks!

Sarah

Sarah Schuett

Manager, Flow Cytometry & Cell Sorting Core

North Carolina State University-CVM

1060 William Moore Drive

Raleigh, NC 27607

919-513-6443

FROM Mara Rocchi

You can try Sigma, 

biorad (exABD serotec)

https://www.bio-rad-antibodies.com/veterinary-antibodies.html

kingfisher

http://www.kingfisherbiotech.com/

hope this helps

Mara Rocchi PhD, MRCVS, MISTR

FACS Manager

Moredun Research Institute

Pentlands Science Park

Bush Loan, Penicuik

EH26 0PZ, Scotland UK

Ph: +44-131-4456128

Fax: +44-131-4456111

e-mail: mara.rocchi@moredun.ac.uk

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FROM Janet Staats

For what it’s worth, I spent most of yesterday researching commercially available canine Abs to build proposed panels for flow-based phenotyping and functional (cytokine) assays.  This was in advance of an upcoming Canine Oncology meeting, to be used as an illustration of what kind of panels we might be able to create for immune monitoring.

I am attaching two refs sent to me by my colleague at the NCI, Kathy McKinnon, as well as three proposed flow panels I created based on information I found yesterday and attached refs.  I have not tested these panels, only configured them as potential options that might work on our LSRFortessa.

Below is a list of manufacturers/distributor that I found with conjugated anti-canine antibodies for flow cytometry.

eBioscience (eB) = ThermoFisher (TF)

Antibodies Online (AO)

BioRad (BR, formerly AbD Serotech)

R&D Systems (RD)

BD Biosciences (BD)

BioLegend (BL)

Clinisciences (CS in France)

Hopefully this will come in handy for you. It would be great if this information could actually be used to create flow-based panels, at least by one of us.

Best regards,

Janet Staats

Scientific/Research Laboratory Manager

Duke University Medical Center

Duke Immune Profiling Core

119 SORF (Bldg 41), 915 S. LaSalle St

Durham, NC 27710-2926

E-mail: janet.staats@duke.edu

Office: 919.684.9807

Lab 1: 919.684.3315

Lab 2: 919-684.3397

Fax: 919.684.4288

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FROM Anne Avery

 Attached is the list of antibodies we use for phenotyping canine lymphoproliferative disease as well as some functional studies (like foxp3).  There are some antibodies to macrophages/monocytes/dendritic cells that are not on the list, which are used by some groups (CD11b, CD11c, CD11d) that we have less familiarity with.

Milliplex and has several canine cytokine bead-based assays which work well with some small exceptions (for example, you cannot detect IFNg in the serum with this assay).  R&D systems has several ELISAs.  We have used the IL-2 ELISA, which works well, and IL-4 which appears (in our hands anyway) has low sensitivity.

Happy to share any unpublished information that we might have about these.  

Anne Avery, VMD, PhD

Associate Professor

Department of Microbiology, Immunology and Pathology

College of Veterinary Medicine and Biomedical Sciences

200 West Lake Street

Fort Collins, CO 80523

970-491-6138 (office)

970-491-1170 (lab)

970-491-4242 (fax)csu-cvmbs.colostate.edu/academics/mip/ci-lab

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