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
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:
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:
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:
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!
Katie Quarfoth
Senior Key Account Manager
614 McKinley Place NE Minneapolis, MN 55413-2610
Office 612 379 6597 Toll Free 800 343 7475
bio-techne.com Katie.Quarfoth@bio-techne.com
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FROM Davis Seelig
As far as cell phenotyping, we've had good success with antibodies from Serotec (another member of the Bio-Rad family). They have a canine section of their website:
https://www.bio-rad-antibodies.com/dog-canine-antibodies.html
Best,
-davis
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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------------------------------------------------------------
FROM Kerry White
I have a few antibodies that I can recommend for immunophenotyping. Below is from an assay I ran in canine whole blood including titrated concentration.
CD3-FITC clone CA17.2A12 BioRad Catalog #MCA1774F used 1:150
CD21-PE clone CA2.1D6 BioRad Catalog #MCA1781.PE used 1:4
CD14-Pacific Blue Clone TUK4 BioRad Catalog # MCA1568PB used 1:100
Hope this helps,
Kerry
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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 managerMicroscopy, Imaging and Cytometry Resources CoreBarbara Ann Karmanos Cancer InstituteWayne State University(313)576-8341 ------------------------------------------------------------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
Antigen
Clone
CD1a
CA9.AG5
CD1a
NA1/34-HLK
CD1b
CC20
CD1c
CA13.9H11
CD3
CA17.2A12 ST
CD3
3-12
CD4
CA13.1E4
CD4
YKIX302.9
CD5
YKIX322.3
CD8 alpha
CA9.JD3
CD8 alpha
YCATE55.9
CD8 beta
CA15.4G2
CD11/18
YKIX490.6.4
CD11a
CA11.4D3
CD11a
HI111
CD11b
CA16.3E10
CD11b
M1/70
CD11c
CA11.6A1
CD11c
BU15
CD11d
CA16.3D3
CD11d
CA11.8H2
CD14
TUK4
CD18
CA1.4E9
CD18
YFC118.3
CD21
CA2.1D6
B cells
B-ly4
CD22
RFB4
CD25
P4A10
CD34
1H6
CD44
IM7
CD45
CA12.10C12
CD45
YKIX716.13
CD45RA
CA4.1D3
CD49d
CA4.5B3
CD61
SZ21
CD79a
HM57
CD79b
AT107-2
CD90
CA1.4G8
CD90
YKIX337.217
CD90
5E10
CD117
ACK45
CD117
YB58B8
MHC II
CA2.1C12
YKIX334.2
Anti-PMN
CADO48A
TCR a/b*
CA15.8G7
TCR g/d*
CA20.6A3
* restricted use - P.F. Moore
***************************************
Dorothee Bienzle, DVM, PhD, Dipl. ACVP
Professor of Veterinary Pathology
Department of Pathobiology
University of Guelph
Room 3822, Building 89
419 Gordon St.
Guelph, ON N1G 2W1
CANADA
Tel 519.824.4120 x54351
Fax 519.824.5930
dbienzle@uoguelph.ca
------------------------------------------------------------ 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
http://www.cvm.ncsu.edu/ccmtr/flocyt.html
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FROM Mara Rocchi
You can try Sigma, biorad (exABD serotec)https://www.bio-rad-antibodies.com/veterinary-antibodies.htmlkingfisherhttp://www.kingfisherbiotech.com/hope this helpsMara Rocchi PhD, MRCVS, MISTRFACS ManagerMoredun Research InstitutePentlands Science ParkBush Loan, PenicuikEH26 0PZ, Scotland UKPh: +44-131-4456128Fax: +44-131-4456111e-mail: mara.rocchi@moredun.ac.uk------------------------------------------------------------
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 StaatsScientific/Research Laboratory ManagerDuke University Medical CenterDuke Immune Profiling Core119 SORF (Bldg 41), 915 S. LaSalle StDurham, NC 27710-2926E-mail: janet.staats@duke.eduOffice: 919.684.9807Lab 1: 919.684.3315Lab 2: 919-684.3397Fax: 919.684.4288------------------------------------------------------------
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, PhDAssociate ProfessorDepartment of Microbiology, Immunology and PathologyCollege of Veterinary Medicine and Biomedical Sciences200 West Lake StreetFort Collins, CO 80523970-491-6138 (office)970-491-1170 (lab)970-491-4242 (fax)csu-cvmbs.colostate.edu/academics/mip/ci-lab------------------------------------------------------------
