Anti-PD-L1 monoclonal antibody with broad species cross-reactivity

Anti-PD-L1, Rat Mono (6C11-3A11)

Cat.# FDV-0054 / Size: 100 µl

Datasheet

The clone 6C11-3A11 is an anti-PD-L1 monoclonal antibody generated using bovine PD-L1 (Programmed cell death 1 ligand 1) as the immunogen. Subsequent analyses have confirmed that it exhibits cross-reactivity across a wide range of animal species, including dog, horse, pig, sheep, goat, and buffalo. It is useful for immunohistochemical staining and flow cytometry analyses.

PD-L1 in various animal species and antibody clone 6C11-3A11

Programmed cell death 1 (PD-1) is a member of the immunoglobulin superfamily expressed mainly on T cells and inhibits T cell proliferation and effector functions. PD-L1, a ligand for PD-1, is known to be induced by various mechanisms, including inflammatory cytokines, and is highly expressed in many types of cancer cells. PD-L1 is attracting attention as an immune escape mechanism for cancer cells. Therefore, inhibition of PD-1/PD-L1 interaction is expected to be a promising anti-tumor strategy, and basic research, drug discovery, and clinical studies are actively being conducted in humans and rodents, and several approved drugs have been launched. On the other hand, the function of PD-L1 in animals such as cattle and dogs has not been well analyzed. PD-L1 has high homology among animal species, suggesting its function may be conserved across species.

The monoclonal antibody 6C11-3A11 against bovine PD-L1 extracellular region has been found to react with PD-L1 in a wide range of animal species. It is expected to be an excellent experimental tool for immunohistochemical staining and flow cytometry analysis. 6C11-3A11 is suitable for the detection and quantification of PD-L1 in these animal species and is being used intensively to elucidate PD-L1 function in each animal. For example, it has been used to observe infection- and inflammation-dependent PD-L1 upregulation in cattle, to detect PD-L1 expression in malignant tumors, and to elucidate the mechanism of PD-L1 expression induction by inflammatory cytokines in dogs, and to analyze stimulus-dependent PD-L1 expression responses in pigs, horses, and sheep. In details, please check ☞ reference.

Features

• Immunogen: Bovine PD-L1 extracellular region

• Host Species: Rat

• Clonality: monoclonal

• Isotype: Rat IgG_2a, κ

• Applications:

*Cow, Dog, Pig, Sheep, Goat, Buffalo

−  Immunohistochemistry (parrafin)

−  Flow cytometry

*Horse

− Neutralizing activity (inhibition of PD-L1 binding to PD-1)

• Specificity：

− Cow, Dog, Horse, Pig, Sheep, Goat, Buffalo

− Not reactive with Cat and Human

Reference

Cow

• Sajiki, Y., et al., "Prostaglandin E₂ induction suppresses the Th1 immune responses in cattle with Johne’s disease.", Infect. Immun., 86(5), e00910～e00917 (2018). [PMID：29483289]

• Sajiki, Y., et al., "Prostaglandin E₂-induced immune exhaustion and enhancement of antiviral effects by anti-PD-L1 antibody combined with COX-2 inhibitor in bovine leukemia virus infection.", J. Immunol., 203(5), 1313～1324 (2019). [PMID：31366713]

• Goto, S., et al., "Upregulation of PD-L1 expression by prostaglandin E₂ and the enhancement of IFN-γ by anti-PD-L1 antibody combined with a COX-2 inhibitor in Mycoplasma bovis infection.", Front. Vet. Sci., 7, 12 (2020). [PMID：32154274]

• Sajiki, Y., et al., "Tick saliva-induced programmed death-1 and PD-ligand 1 and its related host immunosuppression.", Sci. Rep., 11(1), 1063 (2021). [PMID：33441793]

Dog

• Takeuchi, H., et al., "Expression analysis of canine CMTM6 and CMTM4 as potential regulators of the PD-L1 protein in canine cancers.", Front. Vet. Sci., 7, 330 (2020). [PMID：32596272]

• Maekawa, N., et al., "PD-L1 immunohistochemistry for canine cancers and clinical benefit of anti-PD-L1 antibody in dogs with pulmonary metastatic oral malignant melanoma.", NPJ Precis. Oncol., 5(1), 10 (2021). [PMID：33580183]

• Gulay, K.C.M., et al., "Hemangiosarcoma cells induce M2 polarization and PD-L1 expression in macrophages.", Sci. Rep., 12(1), 2124 (2022) . [PMID：35136176]

• Owaki, R., et al., "Regulation of programmed death ligand 1 expression by interferon-γ and tumour necrosis factor-α in canine tumour cell lines.", Vet. Comp. Oncol., 21(2), 279～290 (2023). [PMID：36802270]

Pig

• Ganbaatar, O., et al., "Programmed death-ligand 1 expression in swine chronic infections and enhancement of interleukin-2 production via programmed death-1/programmed death-ligand 1 blockade.", Immun. Inflamm. Dis., 9(4), 1573～1583 (2021). [PMID：34414683]

Horse

• Ganbaatar, O., et al., "PD-L1 expression in equine malignant melanoma and functional effects of PD-L1 blockade.", PLOS One, 15(11), e0234218 (2020). [PMID：33216754]

Sheep

• Tiyamanee, W., et al., "Molecular characterization of immunoinhibitory factors PD-1/PD-L1 in sheep.", Vet. Immunol. Immunopathol., 261, 110609 (2023). [PMID：37201379]

Application example

PD-L1 expression in canine tumor tissues (Immunohistochemical analysis)

Formalin-fixed, paraffin-embedded tissue sections from canine squamous cell carcinoma, transitional cell carcinoma, and diffuse large B-cell lymphoma were subjected to immunohistochemical staining using the antibody 6C11-3A11 (5 μg/mL) and an HRP-conjugated anti-rat IgG secondary antibody. Hematoxylin was used as a counterstain. Staining signals were observed in all tumor tissues treated with the 6C11-3A11 antibody.

Observation of stimulus-dependent induction of PD-L1 expression in canine tumor cell lines (Flow cytometry)

The canine malignant melanoma cell line CMeC was stimulated with various cytokines (TNF-α, IFN-γ and TNF-α/IFN-γ). Cells were then stained with the antibody 6C11-3A11 (10 μg/mL) and an APC-conjugated anti-rat Ig secondary antibody, followed by analysis by flow cytometry. Increased PD-L1 expression was observed upon treatment with either TNF-α or IFN-γ. Moreover, combined treatment with both cytokines induced a greater increase in PD-L1 expression compared with either stimulus alone.

FOR RESEARCH USE ONLY. NOT FOR USE IN HUMANS.

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