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Anti-PD-L1 monoclonal antibody with broad species cross-reactivity Anti-PD-L1, Rat Mono (6C11-3A11)

Date：April 03 2026Web Page No：95045

Funakoshi Co.,Ltd.

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
Features
Application example
Reference
Product information

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.

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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

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Cow

Validation for bovine PD-L1 (Flow cytometry)

CHO cells overexpressing bovine PD-L1 fused with EGFP (PD-L1-EGFP) or EGFP were treated with anti-PD-L1 antibody (6C11-3A11; 10 μg/ml) or rat IgG2a isotype (10 μg/ml) as a negative control for 20 min at R.T. Then, cells were treated with anti-Rat Immunoglobulin (Ig)-allophycocyanin (APC) conjugated for 20 min at R.T. and applied to flow cytometry analysis. While 6C11-3A11 shows little binding to EGFP-expressing cells, 6C11-3A11 significantly binds to bovine PD-L1-EGFP expressing cells.

Bovine PD-L1-EGFP expressing cells		EGFP expressing cells

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	Rat IgG_2a control		6C11-3A11

Detection of endogenous PD-L1 on bovine blood cells (flow cytometry)

Bovine peripheral blood mononuclear cells (PBMCs) and CD14⁺ cells were treated with PBS containing 10% goat serum at 25°C for 20 minutes. The cells were then incubated with 6C11-3A11 (10 μg/mL) or a rat IgG₂a isotype control at 25°C for 20 minutes. Subsequently, the cells were incubated with an APC-conjugated anti-rat Ig secondary antibody at 25°C for 20 minutes and analyzed by flow cytometry. PD-L1 was detected on PBMCs and CD14⁺ monocytes.


Click on the image to enlarge

	Bovine PBMC/Rat IgG_2a control	Bovine CD14⁺ cells/Rat IgG_2a control


	Bovine PBMC/6C11-3A11	Bovine CD14⁺ cells/6C11-3A11

PD-L1 expression in cattle with Johne’s disease (Immunohistochemical analysis)

Sections of ileal tissue (formalin-fixed, paraffin-embedded) from healthy cattle and cattle with Johne’s disease were prepared and subjected to immunohistochemical staining using 6C11-3A11 (10 μg/mL), an HRP-conjugated anti-rat IgG secondary antibody, and DAB. Hematoxylin was used as a counterstain. Marked PD-L1 expression was observed in the lesional ileal tissue of cattle with Johne’s disease.


Uninfected（normal ileum）		Johne's disease（ileum）

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M：M. avium subsp. Paratiberculosis infected macrophages
▲：M. avium subsp. Paratiberculosis infected Langhans giant cells

PD-L1 expression in bovine skin tissue at tick bite sites (Immunohistochemical analysis)

Sections of skin tissue (formalin-fixed, paraffin-embedded) derived from normal cattle and cattle infested with the cattle tick (Rhipicephalus microplus) were prepared. Immunohistochemical staining was performed using 6C11-3A11 (10 μg/mL), an HRP-conjugated anti-rat IgG secondary antibody, and DAB. Hematoxylin staining was used as a counterstain (◀ indicates Rhipicephalus microplus). An increased number of PD-L1–positive cells was observed in cattle infested with the cattle tick.

Tick-nonattached site		Tick-attached site

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Dog

Evaluation of reactivity against canine PD-L1 (Flow cytometry)

CHO-DG44 cells stably expressing canine PD-L1–EGFP or EGFP alone were incubated with 6C11-3A11 (10 μg/mL) for 30 minutes at room temperature, followed by incubation with an allophycocyanin (APC)-conjugated anti-rat Ig secondary antibody for 30 minutes. The antibody-treated cells were then analyzed by flow cytometry. A marked increase in fluorescence signal was observed in canine PD-L1–EGFP–expressing cells upon staining with 6C11-3A11.

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.

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.

	Squamous cell carcinoma	Transitional cell carcinoma	Diffuse large B-cell lymphoma
6C11-3A11

2^nd antibody only

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

Immunofluorescence staining was performed on tissue sections of canine hemangiosarcoma (HSA) using the antibody 6C11-3A11 (10 μg/mL) and an anti-Iba1 antibody (macrophage marker). PD-L1 signals were observed to partially colocalize with macrophages.

Anti-PD-L1	Anti-Iba1	Merge

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Pig

Evaluation of reactivity against Porcine PD-L1 and analysis of stimulus-dependent hanges in PD-L1 Expression Levels (Flow cytometry)

Evaluation of reactivity to porcine PD-L1		Analysis of stimulus-dependent changes in PD-L1 expression

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	EGFP/Rat IgG_2a control		Unstimulated swine PBMC/Rat IgG_2a control


	EGFP/6C11-3A11		Unstimulated swine PBMC/6C11-3A11


	Swine PD-L1-EGFP/Rat IgG_2a control		Swine PBMC stimulated with IFN-γ/Rat IgG_2a control


	Swine PD-L1-EGFP/6C11-3A11		Swine PBMC stimulated with IFN-γ/6C11-3A11

COS-7 cells expressing porcine PD-L1-EGFP were incubated with 6C11-3A11 (10 μg/mL) at 25 ℃ for 20 minutes, followed by incubation with allophycocyanin (APC)-conjugated anti-rat Ig secondary antibody at 25  ℃ for 20 minutes. The antibody-treated cells were subsequently analyzed by flow cytometry. In cells expressing porcine PD-L1-EGFP, 6C11-3A11 staining resulted in a marked increase in fluorescent signal.		Porcine peripheral blood mononuclear cells (PBMCs) were cultured at 37  ℃ with 5% CO₂ for 24 hours in the presence or absence of porcine IFN-γ (10 μg/mL). The cultured cells were blocked with PBS containing 10% goat serum at 25  ℃ for 20 minutes, followed by incubation with 6C11-3A11 (10 μg/mL) at 25 °C for 20 minutes, and then with APC-conjugated anti-rat Ig secondary antibody at 25  ℃ for 20 minutes. The cells were subsequently analyzed by flow cytometry. IFN-γstimulation resulted in a marked increase in PD-L1 expression on PBMCs.

PD-L1 Expression in Pigs with Chronic Infection (Immunohistochemical analysis)

Formalin-fixed, paraffin-embedded lung tissue sections from healthy pigs and pigs infected with porcine reproductive and respiratory syndrome virus (PRRSV) or Mycoplasma hyopneumoniae were subjected to immunohistochemical staining using the 6C11-3A11 antibody (10 μg/mL) and HRP-conjugated anti-rat IgG secondary antibody, with hematoxylin used for counterstaining. As a negative control, lung tissue sections from healthy pigs were stained with the HRP-conjugated anti-rat IgG secondary antibody alone. In infected tissues, staining signals were observed upon treatment with the 6C11-3A11 antibody.

Uninfected	PRRSV-infected	M. hyopeneumoniae-infected

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Horse

Evaluation of reactivity to equine PD-L1 and analysis of stimulus-dependent changes in PD-L1 expression (Flow cytometry)

Evaluation of reactivity to equine PD-L1		Analysis of stimulus-dependent changes in PD-L1 expression

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	EGFP/Rat IgG_2a control		Equine fresh PBMC/Rat IgG_2a control	Equine PBMC stimulated with IFNγ/Rat IgG_2a control


	EGFP/6C11-3A11		Equine fresh PBMC/6C11-3A11	Equine PBMC stimulated with IFNγ/6C11-3A11


	Equine PD-L1-EGFP/Rat IgG_2a control


	Equine PD-L1-EGFP/6C11-3A11

COS-7 cells expressing equine PD-L1-EGFP were incubated with 6C11-3A11 (10 μg/mL) at 25 °C for 20 minutes, followed by incubation with allophycocyanin (APC)-conjugated anti-rat Ig secondary antibody at 25 °C for 20 minutes. The antibody-treated cells were then analyzed by flow cytometry. In cells expressing equine PD-L1-EGFP, 6C11-3A11 staining resulted in a marked increase in fluorescent signal.		Equine peripheral blood mononuclear cells (PBMCs) were cultured at 37 °C with 5% CO₂ for 24 hours in the presence or absence of IFN-γ (10 μg/mL). The cultured cells were blocked with PBS containing 10% goat serum at room temperature for 20 minutes, followed by incubation with 6C11-3A11 (10 μg/mL) at room temperature for 20 minutes, and then with APC-conjugated anti-rat Ig secondary antibody at room temperature for 20 minutes. The cells were subsequently analyzed by flow cytometry. IFN-γ stimulation resulted in a marked increase in PD-L1 expression on PBMCs.

PD-L1 Expression in Equine Melanoma Tissue (Immunohistochemical analysis)

Formalin-fixed, paraffin-embedded tissue sections of equine cutaneous melanoma were subjected to immunohistochemical staining using the 6C11-3A11 antibody (10 μg/mL) and HRP-conjugated anti-rat IgG secondary antibody, with hematoxylin used for counterstaining. As a negative control, sections were stained with the HRP-conjugated anti-rat IgG secondary antibody alone. Strong staining signals were observed in equine melanoma tissues upon treatment with the 6C11-3A11 antibody.

Control		PD-L1（6C11-3A11）

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Sheep

Evaluation of reactivity to ovine PD-L1 and analysis of stimulus-dependent changes in PD-L1 expression (Flow cytometry)

Evaluation of reactivity to ovine PD-L1		Analysis of stimulus-dependent changes in PD-L1 expression

Click on the image to enlarge

	EGFP/Rat IgG_2a control		Unstimulated ovine PBMC/Rat IgG_2a control


	EGFP/6C11-3A11		Unstimulated ovine PBMC/6C11-3A11


	Ovine PD-L1-EGFP/Rat IgG_2a control		Ovine PBMC stimulated with PWM/Rat IgG_2a control


	Ovine PD-L1-EGFP/6C11-3A11		Ovine PBMC stimulated with PWM/6C11-3A11

COS-7 cells expressing ovine PD-L1-EGFP were incubated with 6C11-3A11 (10 μg/mL) at room temperature for 20 minutes, followed by incubation with allophycocyanin (APC)-conjugated anti-rat Ig secondary antibody at room temperature for 20 minutes. The antibody-treated cells were then analyzed by flow cytometry. In cells expressing ovine PD-L1-EGFP, 6C11-3A11 staining resulted in a marked increase in fluorescent signal.		Ovine peripheral blood mononuclear cells (PBMCs) were cultured at 37 °C with 5% CO₂ for 24 hours in the presence or absence of pokeweed mitogen (PWM, 10 μg/mL). The cultured cells were blocked with PBS containing 10% goat serum at room temperature for 20 minutes, followed by incubation with 6C11-3A11 (10 μg/mL) at room temperature for 30 minutes. The cells were then incubated with APC-conjugated anti-rat Ig secondary antibody at room temperature for 30 minutes and analyzed by flow cytometry. PWM stimulation resulted in a marked increase in PD-L1 expression on PBMCs.

Analysis of PD-L1 Expression in Ovine Tissues Infected with Listeria monocytogenes(Immunohistochemical analysis)

Formalin-fixed, paraffin-embedded brain tissue sections from sheep infected with Listeria monocytogenes were subjected to immunohistochemical staining using the 6C11-3A11 antibody (10 μg/mL), HRP-conjugated anti-rat IgG secondary antibody, and DAB. Strong staining signals were observed in macrophages (white arrows) present in the brain tissue of L. monocytogenes-infected sheep.

Listeria monocytogenesに感染したヒツジにおけるPD-L1発現解析（免疫組織化学染色）

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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]

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Product information

[Date : April 11 2026 00:08]

Detail Product Name Product Code Supplier Size Price

Anti-PD-L1, Rat-Mono(6C11-3A11)
DatasheetThis may not be the latest data sheet. FDV-0054 FNAFunakoshi Co.,Ltd. 100 µl $500

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Description	Rat monoclonal antibody (clone name 6 C11-3 A11) established using the extracellular region of bovine PD-L1 as an antigen. It has been confirmed that it reacts with PD-L1 of not only cattle but also dog, horse, pig, sheep and buffalo. It can be used for immunostaining and flow cytometry.
Storage	-20°C
Antigen Species	Bovine	Host Species	Rat	Class	IgG	Label	Unlabeled
Cross Reactivity	Cow/Dog/Horse/Pig/Sheep/Goat/Buffalo			Application	FCM,IHC
Clonality		Format		Purification	Purified	Absorption
Link	Cancer-Related Antibody

[Date : April 11 2026 00:08]

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

DatasheetThis may not be the latest data sheet.

Product Code: FDV-0054
Supplier: FNA
Size: 100µl
Price: $500

Description	Rat monoclonal antibody (clone name 6 C11-3 A11) established using the extracellular region of bovine PD-L1 as an antigen. It has been confirmed that it reacts with PD-L1 of not only cattle but also dog, horse, pig, sheep and buffalo. It can be used for immunostaining and flow cytometry.
Storage	-20°C
Antigen Species	Bovine	Host Species	Rat
Class	IgG	Label	Unlabeled
Cross Reactivity	Cow/Dog/Horse/Pig/Sheep/Goat/Buffalo	Application	FCM,IHC
Clonality		Format
Purification	Purified	Absorption
Link	Cancer-Related Antibody

Contact

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