A listing for canine specific reagents that have been suggested and, in come cases, validated by the National Cancer Institute (NCI) working group for canine immunotherapeutics. This is a resource for others that might wish to study canine cancers and other immunologic issues.
- Reagents Panels
- Basic B and T cell quantification panel
- B cell panel
- Regulatory T cells
- T cell activation panel
- Alternative T cell function/activation panel (UAB)
- Memory T cell subsets
- T cell exhaustion panel
- Macrophage/DC panel
- Macrophage/DC activation panel
- Myeloid panel
- Myeloid-Derived Suppressor Cells (Supplied by Dr. Garden’s lab)
- NK cell panel
- Macrophage Polarization Panel
- Antibodies
Reagents Panels
Basic B and T cell quantification panel
Last updated: November 22, 2021
Reagents Contact Form
| Antigen | Clone | Fluorochrome | Supplier | Host Species | Target species | Isotype |
|---|---|---|---|---|---|---|
| CD3 | CA17.2A12 | FITC | BioRad | Mouse | Dog | IgG1 |
| 1CD5 | YKIX322.2 | PE | BioRad | Rat | Dog | IgG2a |
| CD4 | YKIX302.9 | APC, AF488,AF647, FITC, PB, PE | BioRad | Rat | Dog | IgG2a |
| CD8a | YCATE55.9 | AF647, AF700,FITC,PB,PE | BioRad | Rat | Dog | IgG1 |
| CD21 | CA2.1D6 | AF647,PE, | BioRad | Mouse | Dog | IgG1 |
| 2CD79b | AT107-2 | FITC | BioRad | Rat | Mouse (Dog) | IgG1 |
| 3CD79a | Hm47 | PE, APC, PerCP-Cy5.5 | ThermoFisher | Mouse | Human | IgG1k |
| Live/ dead | L34966 Cat.no. 65-0865 (fixable, Ex 633nm/Em780) | ThermoFisher | ||||
| 4 | ||||||
| CD45 | YKIX716.13 (or CA12.10C12) | APC, AF488,AF647 FITC, biotin, PE | BioRad | Rat | Dog | IgG2b |
1CD5 yields more stable and reliable flow output than CD3 in Mason Lab and Garden Lab;
2CD79b is used by Garden Lab for labeling canine B cells (intracellular staining);
3CD79b is used by Garden Lab for labeling canine B cells (intracellular staining);
B cell panel
Last updated: November 22, 2021
Reagents Contact Form
| Antigen | Clone | Fluorochrome | Supplier | Host Species | Target species | Isotype |
|---|---|---|---|---|---|---|
| CD21 | CA2.1D6 | AF647,PE, | BioRad | Mouse | Dog | IgG1 |
| 6Iso Ctrl CD21 | Unspecified(Cat.MCA928A647) | AF647 | BioRad | Mouse | IgG1 | |
| CD79b | AT107-2 | FITC | BioRad | Rat | Mouse (Dog) | IgG1 |
| 7Iso Ctrl CD79b | Unspecified(Cat.MCA1211F) | FITC | BioRad | Rat | IgG1 | |
| MHCII | YCATE55.9 | AF647, AF700,FITC,PB,PE | BioRad | Rat | Dog | IgG1 |
| Surface IgG | Polyclonal | FITC | Origene | Sheep | Dog | Polyclonal |
6,7Isotype controls are not essential for lymphocytes, could be replaced by FMO controls
Regulatory T cells
Last updated: November 22, 2021
Reagents Contact Form
| Antigen | Clone | Fluorochrome | Supplier | Host Species | Target species | Isotype |
|---|---|---|---|---|---|---|
| CD5 | YKIX322.2 | PE | BioRad | Rat | Dog | IgG2a |
| CD4 | YKIX302.9 | APC, AF488,AF647, FITC, PB, PEPE-Cy7 | BioRad, ThermoFisher | Rat | Dog | IgG2a |
| CD25 | P4A10 | FITC, PE | BioRad | Mouse | Dog | IgG1 |
| Live/ dead | L34966 Cat.no. 65-0865 (fixable, Ex 633nm/Em780) | ThermoFisher | ||||
| FOXP3 | FJK-16S | AF488, FITC, PE, APC | ThermoFisher | Rat | Mouse | IgG2a Kappa |
| Iso Ctrl FoxP3 | eBR2a | APC | ThermoFisher | Rat | IgG2a Kappa | |
| 4Helios | 22F6 | PE, APC, eFluor710, eFluor450, PE-Cy7, APC-eFluor780 | ThermoFisher | Armenian Hamster | Human/Mouse (Dog) | IgG |
| Iso Ctrl Helios | eBio299Arm | APC | ThermoFisher | Armenian Hamster | IgG | |
| CD45 | YKIX716.13 (or CA12.10C12) | APC, AF488,AF647 FITC, biotin, PE, eFluor450 | BioRad, ThermoFisher | Rat | Dog | IgG2b |
| CTLA4 5(developed for WB) | 3 Mouse monoclonals mixed | Unconjugated | Abmart (X3-Q9XSI1 [ABX]) | Mouse | Dog | Monoclonal, 3 combined IgG |
4 Helios was proposed for labeling thymic Tregs but currently remains controversial
5 Might considering manually conjugate for flow use but expensive (http://www.ab-mart.com/dog/anti-CTLA4%20(Q9XSI1)_X3.html)
T cell activation panel
Last updated: November 22, 2021
Reagents Contact Form
| Antigen | Clone | Fluorochrome | Supplier | Host Species | Target species | Isotype |
|---|---|---|---|---|---|---|
| CD5 | YKIX322.2 | PE | BioRad | Rat | Dog | IgG2a |
| CD25 | P4A10 | FITC, PE | BioRad | Mouse | Dog | IgG1 |
| MHCII | YKIX334.2 | APC, FITC | ThermoFisher | Rat | Dog | IgG2a kappa |
| Ki67 | 20Raj1 | PE-Cy7 | Invitrogen | Mouse | Human | IgG1k |
| Granzyme B | GB11 | PE, FITC | BD Pharmingen, Biolegend | Mouse | Human | IgG1k |
| IFN-g | CC302 | FITC, PE, Biotin, AF-488, AF-647 | BioRad | Mouse | Bovine | IgG1 |
| TNF-a | Mab11 | FITC, AF488, PE | ThermoFisher | Mouse | Human | IgG1k |
| IL-4 | CC303 | FITC, PE | BioRad | Mouse | Bovine | IgG2a |
| CD107b | AC17 | FITC, PE, AF-647 | BioRad | Mouse | Dog | IgG1 |
| Live/ dead | L34966 | ThermoFisher |
Alternative T cell function/activation panel (UAB)
Last updated: November 22, 2021
Reagents Contact Form
| Antigen | Clone | Fluorochrome | Supplier | Host Species | Target species | Isotype |
|---|---|---|---|---|---|---|
| CD3 | CA17.2A12 | FITC | BioRad | Mouse | Dog | Ig1 |
| IL-4 | CC303 | FITC, PE | BioRad | Mouse | Bovine | IgG2a |
| CD4 | YKIX302.9 | APC, AF488,AF647, FITC, PB, PE | BioRad | Rat | Dog | IgG2a |
| IFN-g | CC302 | FITC, PE, Biotin, AF-488, AF-647 | BioRad | Mouse | Bovine | IgG1 |
| CD8a | YCATE55.9 | AF647, AF700,FITC,PB,PE | BioRad | Rat | Dog | IgG1 |
| FOXP3 | FJK-16S | AF488, FITC, PE | ThermoFisher | Rat | Mouse | IgG2a Kappa |
| Live/ dead | L34966 | ThermoFisher |
Memory T cell subsets
Last updated: November 22, 2021
Reagents Contact Form
| Antigen | Clone | Fluorochrome | Supplier | Host Species | Target species | Isotype |
|---|---|---|---|---|---|---|
| CD3 | CD3-12 | PB | BioRad | Rat | Human | ? |
| CD4 | YkIX302.9 | PE-Cy7 | eBioscience | Rat | Dog | IgG2a kappa |
| CD4 | YKIX302.9 | APC, AF488,AF647, FITC, PB, PEPE-Cy7 | BioRad, ThermoFisher | Rat | Dog | IgG2a |
| CD8a | YCATE55.9 | AF647, AF700,FITC,PB,PE | BioRad | Rat | Dog | IgG1 |
| CD28 | 5B8 | APC | eBioscience | Mouse | Dog | IgG1 kappa |
| CD44 | YKIX337.8 | FITC | ThermoFisher | Rat | Dog | IgG2a kappa |
| CD62L | FMC46 | PE | BioRad | Mouse | Human | IgG2b |
| CD45RA | CA4.1D3 | Biotin | BioRad | Mouse | Dog | IgG1 |
| +/-CCR7 | 150503 | PE, AF-647, AF-700, BV421, FITC, PerCP-Cy5.5, V450, | BD | Mouse | Human | IgG2a |
| Live/ dead | L34966 | ThermoFisher |
1. Withers & Rebhun Multi-color flow cytometry for evaluating age related changes in memory lymphocyte subsets in dogs. Developmental and Comparative Immunology
T cell exhaustion panel
Last updated: November 22, 2021
Reagents Contact Form
| Antigen | Clone | Fluorochrome | Supplier | Host Species | Target species | Isotype |
|---|---|---|---|---|---|---|
| CD5 | YKIX322.2 | PE | BioRad | Rat | Dog | IgG2a |
| PD-1 | 3B6 | |||||
| CD28 | 5B8 | Unconjugated | ||||
| Ki67 | 20Raj1 | PE-Cy7 | Invitrogen | Mouse | Human | IgG1k |
| EOMES | WD1928 | PE-Cy7 | Invitrogen | Mouse | Human | IgG1k |
| Mouse IgG | AF-488 | Donkey | Mouse |
Macrophage/DC panel
Last updated: November 22, 2021
Reagents Contact Form
| Antigen | Clone | Fluorochrome | Supplier | Host Species | Target species | Isotype |
|---|---|---|---|---|---|---|
| CD11b | CA16.3E10 | Unconjugated | AbDSerotec | Mouse | Dog | IgG1 |
| CD11c | CA11.6A1 | Unconjugated | BioRad | Mouse | Dog | IgG1 |
| CD14 | TUK4 | AF-647 | AbDSerotec | Mouse | Human | |
| MHCII | YKIX334.2 | APC, FITC | ThermoFisher, AbD Serotec | Rat | Dog | IgG2a kappa |
| CD4 | YKIX302.9 | APC, AF488,AF647, FITC, PB, PE | BioRad | Rat | Dog | IgG2a |
| CCR2 | ||||||
| CD80 |
Macrophage/DC activation panel
Last updated: November 22, 2021
Reagents Contact Form
| Antigen | Clone | Fluorochrome | Supplier | Host Species | Target species | Isotype |
|---|---|---|---|---|---|---|
| CD11b | CA16.3E10 | Unconjugated | AbDSerotec | Mouse | Dog | IgG1 |
| CD86 | BU63* | PE | ? | Mouse | Human | IgG1 |
| CD40 | LOB7/6* | FITC | ? | Mouse | Human | IgG2a |
| MHCII | YKIX334.2 | APC, FITC | ThermoFisher, AbD Serotec | Rat | Dog | IgG2a kappa |
| CD18 | CA1.4E9 | AF-647 | BioRad | Mouse | Canine | IgG1 |
| TNF-a | Mab11 | FITC, AF488, PE | ThermoFisher | Mouse | Human | IgG1k |
| MCP-1 | Polyclonal | Rabbit | Pig |
Myeloid panel
Last updated: November 22, 2021
Reagents Contact Form
| Antigen | Clone | Fluorochrome | Supplier | Host Species | Target species | Isotype |
|---|---|---|---|---|---|---|
| HLA-DR | ? | ? | ? | ? | ? | ? |
| CD45 | ||||||
| CD11b | CA16.3E10 | Unconjugated | AbDSerotec | Mouse | Dog | IgG1 |
| Macrophage | MAC387 | FITC, PE, ALC, AF-488 | Biorad | Mouse | Human | IgG1 |
| MHCII | YKIX334.2 | APC, FITC | ThermoFisher, AbD Serotec | Rat | Dog | IgG2a kappa |
| CD14 | TUK4 | AF-647 | AbDSerotec | Mouse | Human | |
| Neutrophils 8(Ly6G/C) | RB6-8C5 | FITC, PE, AF488 | ThermoFisher | Rat | Mouse | IgG2b |
| Myleoid cells | CADO48A | FITC | Washington State | Mouse | Dog | IgG1 |
| Live/ dead | L34966 | ThermoFisher |
8 Failedto yield positive staining on canine MDSC cells in Garden Lab
Myeloid-Derived Suppressor Cells (Supplied by Dr. Garden’s lab)
Last updated: November 22, 2021
Reagents Contact Form
| Antigen | Clone | Fluorochrome | Supplier | Host Species | Target species | Isotype |
|---|---|---|---|---|---|---|
| CD11b | M1/70 | AF488 | eBioscience, ThermoFisher | Rat | Mouse | IgG2b |
| MHC II | YKIX334.2 | APC | eBioscience, ThermoFisher | Rat | Dog | IgG2a |
| CD5 | YKIX322.3 | RPE, also available in FITC, APC, AF488 and AF647 | Bio-Rad | Rat | Dog | IgG2a |
| CD21 | CA2.1D6 | RPE, also available in AF647 | Bio-Rad | Mouse | Dog | IgG1 |
| DAPI | BioLegend | |||||
| CADO48A | CADO48A | Unconjugated | Washington State University | Dog | IgG1 | |
| CD14 | TUK4 | RPE-AF647, also available in RPE-AF750, AF700, AF647, AF488, Biotin, FITC, Pacific Blue, Low endodoxin | Bio-Rad | Mouse | Human | IgG2a |
NK cell panel
Last updated: November 22, 2021
Reagents Contact Form
| Antigen | Clone | Fluorochrome | Supplier | Host Species | Target species | Isotype |
|---|---|---|---|---|---|---|
| CD5 | YKIX322.2 | PE | BioRad | Rat | Dog | IgG2a |
| CD8a | YCATE55.9 | AF647, AF700,FITC,PB,PE | BioRad | Rat | Dog | IgG1 |
| NKp46 | 48A | Unconjugated | Jennifer Foltz (NWChildrens) | Mouse | Dog | IgG2a |
| CD94 | HP-3D9 (KLRD1) | APC, FITC, PA, PerCP-Cy5.5, | BDBiosciences | Mouse | Human | IgG1k |
| Live/ dead | L34966 | ThermoFisher |
Macrophage Polarization Panel
Macrophage Polarization Contact Form
| Marker | Localization (ICC or flow) | Clone | Vendor (cat #) |
|---|---|---|---|
| iNOS (M1) | intracellular ICC+flow | Polyclonal | Thermo fisher (#PA3-030A) |
| Arginase (M2) | intracellular ICC+flow | 19/Arginase1 | BD (#610708) |
| CD206 (M2) | plasma membrane + trans memb. (ICC; intracellular staining) | 3.29B1.10 | Beckman Coulter (#IM2741) |
| TGM2 (M2) | Plasma membrane + Cytosol(ICC+flow) | Polyclonal | Bio-Rad (#AHP2532) |
| SOCS1 (M2) | intracellular ICC+flow | Polyclonal | Bio-Rad (#AHP1205) |
Antibodies
| Antigen | Antibody (clone) | Vendor | Antigen retrieval |
|---|---|---|---|
| CD3 | Rabbit polyclonal (anti human) | Dako | Citrate HIER |
| CD79a | Mouse (HM57) | Dako | EDTA HIER |
| MHCII | Mouse (TAL. 1B5) | Dako | Citrate HIER |
| CD204 | SRA-E5 | TransGenics Inc. | Citrate HIER |
| Factor VIII-RA | Rabbit polyclonal | Dako | Proteinase K |
| Lysozyme | Rabbit polyclonal | Dako | Proteinase K |
| MUM1 | Mouse (MUM1p) | Dako | EDTA HIER |
| Myeloid/histiocyte antigen (MAC 387) | Mouse (MAC387) | Dako | Proteinase K |
| CD31 | Mouse (JC/70A) | Dako | Citrate HIER |
| CD18 | Mouse (CA16.3C10) | Peter Moore (UC Davis) | Citrate HIER |
| CD11d | Mouse (CA18.3C6) | Peter Moore (UC Davis) | Citrate HIER |
| CD45 | Mouse (CA12.10C12) | Peter Moore (UC Davis) | Citrate HIER |
| Ki67 | Monoclonal Mouse (Mib-1) | Dako | Citrate HIER |
| FoxP3 | Mouse (ebio7979) | Thermo/eBioscience | Citrate HIER |
| CCL2 | Rabbit polyclonal (ab9669) | Abcam | Citrate HIER |
| Oct-3/4 | Mouse (C-10) | Santa Cruz | Citrate HIER |
| CD117 | Rabbit polyclonal | Dako | Citrate HIER |
| CD163 | Mouse (AM-3K) | TransGenics Inc. | Citrate HIER |
| Pax5 | Mouse (clone 24/Pax5) | BD Biosciences | Citrate HIER |
| Granzyme B | Rabbit polyclonal | Spring Bioscience | Citrate HIER |