PerCP/Cyanine5.5 anti-mouse CD366 (Tim-3) 134012 from BioLegend

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Proceedings of the National Academy of Sciences of the United States of America
July 30 2024

Fig 1: Proton RT increases CAR T trafficking into PDAC tumors through increased MSLN expression. (A) Schematic of in vivo mouse experiments to evaluate the expression of MSLN. 5 × 105 PDA7940b cells were injected S.C. into the right flank of C57BL/6 mice. Tumors were grown until sizes reached ~50 mm3 before treatment with proton RT. Mice were killed, and tumors were harvested for analysis 3- and 8-d post-proton RT. (B) A representative fluorescent microscopy image shows blood vessels (CD31; red) and MSLN (green) of irradiated and unirradiated (NR) PDAC tumors (10× magnification). Scale bar, 100 µm. (C–E) MFI of MSLN levels, as assessed by flow cytometry of tumor single-cell suspension, at (C) day 3 and (D) day 8 post 12 Gy proton RT, as well as (E) the percent of MSLN-positive transcripts detected by RNA-ISH in formalin-fixed paraffin-embedded tumor tissues. Average ± SD and individual values are shown. The Mann–Whitney two-tailed test was used for statistical analysis. (F) Schematic of mouse experiments to evaluate CAR T cell trafficking into tumors. 5 × 105 PDA7940b cells were injected S.C. into the right flank of C57BL/6 mice. Tumors were grown until sizes reached ~50 mm3 before treatment with proton RT, followed by single 120 mg/kg cyclophosphamide administration and tail vein injection of 5 × 106 MSLN-CAR T cells at day two post-RT. Eight days post-RT, mice were killed, and tumors were harvested for analysis. (G) CAR T cell count in tumors assessed by flow cytometry. (H) Representative image from RNA-ISH against CD3 (red), CAR T (pink), and MSLN (green). Scale bar, 50 µm. (I–K) Rates of TIM-3 (I), LAG-3 (J), and PD-1 (K) positive CAR T cells 8 d after 12 Gy proton RT (n = 5 per group). The Mann–Whitney U test was used for statistical analysis. (L and M) Immune cell populations in the TME. (L) M1 macrophages (CD45+F4/80+CD86+MHCII+) at 3 d and (M) MDSC (monocytic) (CD45+CD11B+Ly-6G−Ly-6C+) at 8 d following proton RT. Average ± SD are shown. Kruskal–Wallis one-way ANOVA for multiple comparisons was used for statistical analysis. For all panels in this figure, * indicates P ≤ 0.05, ** indicates P ≤ 0.01, and ns (nonsignificant) indicates P > 0.05.
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J Immunother Cancer
April 02 2025

Fig 2: PSCA-targeted DVD anti-CTLA-4 induces antigen-specific responses and improved T-cell fitness in systemically implanted PSCA+ tumors. (A) Percentage of CD8+ T cells within the immune CD45+ compartment from flow cytometry data from lung tumors of mice (n=3) harvested on day 15 after tumor injection. As in the rest of the figure, representative data from n=1 of two experiments is shown. (B) Percentage of intratumoral FoxP3+ cells within gated CD4+ T cells. (C) Percentage of intratumoral CD44+ CD62L− cells within gated CD8+ T cells. (D) Percentage of intratumoral MC38 antigen-specific ADPGK-reactive CD8+ T cells. (E) Representative flow cytometry pseudocolor dot plots showing MC38 antigen ADPGK reactivity in gated CD8+ T cells from mouse lung tumors. Shown values are percentages within all CD8+. (F) MFI of effector molecule perforin (PRF1) in intratumoral ADPGK+ CD8+ T cells. (G) MFI of proliferation marker Ki67 in gated intratumoral ADPGK+ CD8+ T cells. (H) Percentage of PD-1+ TCF1+ cells in gated intratumoral ADPGK+ T cells. (I) Part-of-whole pie charts showing mean percentages of cells expressing 0, any given 1, any combination of 2, or 3 exhaustion markers PD-1, LAG-3, TIM3 in intratumoral ADPGK+ CD8+ T cells, as assessed by Boolean gating from flow cytometry data. Right, percentage of intratumoral PD-1+ TIM3+ LAG-3+ in ADPGK+ CD8+ T cells. aCTLA-4, anti-cytotoxic T-lymphocyte associated protein 4; DVD, dual variable domain; EV, empty vector; LAG-3, lymphocyte-activation gene 3; MFI, mean fluorescence intensity; PD-1, programmed cell death protein 1; PSCA, prostate stem cell antigen; sDVD, symmetric DVD; unDVD, uncleavable DVD; TIM3, T-cell immunoglobulin and mucin-domain containing-3.
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J Immunother Cancer
April 02 2025

Fig 3: Symmetric DVD induces systemic immune responses. (A) Percentage of CD4+ T cells within the immune CD45+ compartment from flow cytometry data from spleens of mice (n=3) harvested on day 15 after tumor injection. As in the rest of the figure, representative data from n=1 of two experiments is shown. (B) Percentage of FoxP3+ cells within gated CD4+ T cells. (C) Percentage of CD44+ CD62L− cells within gated CD4+ T cells. (D) Percentage of splenic CD8+ T cells within the immune CD45+ compartment. (E) Percentage of splenic MC38 antigen-specific ADPGK-reactive CD8+ T cells. Shown p values obtained by one-way analysis of variance adjusted for multiple comparisons; *p<0.05 versus rest. (F) Representative flow cytometry pseudocolor dot plots showing MC38 antigen ADPGK reactivity in gated CD8+ T cells from mouse spleens. Shown values are percentages within all CD8+. (G) MFI of effector molecule perforin (PRF1) in gated splenic ADPGK+ CD8+ T cells. *p<0.05 versus rest. (H) MFI of proliferation marker Ki67 in gated splenic ADPGK+ CD8+ T cells. *p<0.05 versus rest. (I) Percentage of PD-1+ TCF1+ cells in gated splenic ADPGK+ CD8+ T cells. *p<0.05 versus rest. (J) Part-of-whole pie charts showing mean percentages of cells expressing 0, any given 1, any combination of 2, or 3 exhaustion markers PD-1, LAG-3, TIM3 in splenic ADPGK+ CD8+ T cells, as assessed by Boolean gating from flow cytometry data. Right, percentage of splenic PD-1− TIM3− LAG-3− in ADPGK+ CD8+ T cells. (K) Percentage of CD8+ T cells within the immune CD45+ compartment in mouse tumor-draining lymph nodes (TDLNs). (L) Percentage of MC38 antigen-specific ADPGK-reactive CD8+ T cells in TDLNs. (M) Representative flow cytometry pseudocolor dotplots showing MC38 antigen ADPGK reactivity in gated CD8+ T cells from mouse TDLNs. Shown values are percentages within all CD8+. aCTLA-4, anti-cytotoxic T-lymphocyte associated protein 4; DVD, dual variable domain; EV, empty vector; LAG-3, lymphocyte-activation gene 3; MFI, mean fluorescence intensity; PD-1, programmed cell death protein 1; PSCA, prostate stem cell antigen; sDVD, symmetric DVD; unDVD, uncleavable DVD; TIM3, T-cell immunoglobulin and mucin-domain containing-3.
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Scientific reports
May 30 2025

Fig 4: Flow cytometry analysis in placenta derived from B6-Lgals9tm1glp (KO) and wild type (WT) pregnant mice. (A) Percentage of CD45+ CD3− CD49b+ conventional NK (cNK) cells in total leukocytes. (B) Percentage of CD45+ CD3− CD49b− CD49a+ trNK (tissue-resident NK) cells in total leukocytes. (C) Percentage of CD45+ CD3− CD49b− CD49a+ C366+ (Tim-3+) trNK cells. Tim-3, T-cell immunoglobulin domain and mucin domain-containing molecule-3. (D) Percentage of Zombie− Annexin+ early apoptotic trNK cells in Tim-3 + trNK cells. (E) mRNA expression of Cd244, Havcr2 (Tim-3), Eomes, Tnfa, Il6, and Il10. Data shown as mean ± SD and analyzed by one-way ANOVA with Tukey test (*p < 0.05). N = 4–6 in each group. SFD, standard fat diet, 10% fat; HFD, high fat diet, 60% fat.
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JCI Insight
February 23 2026

Fig 5: IL-21 combined with radiation reshapes the TME and increases the infiltration of CD8+ T cells.(A) Schematic diagram for scRNA-Seq of CD45+ immune cells sorted from MC38 tumors of IR group and IL-21+IR group 9–10 days after radiation. (B) t-SNE plot showing sorted CD45+ immune cells. (C) Nightingale rose chart and heatmap showing the proportion of different subtypes of CD45+ immune cells. (D) Violin plots showing Il21r expression in the different subtypes of CD45+ immune cells. (E) Tumor growth of MC38 tumors subjected to radiation and/or IL-21 with or without anti-CD8 depletion antibody. (F) t-SNE plot showing the subtypes of CD8+ T cells. (G) Pseudo-time analysis of the subtypes of CD8+ T cells. (H) Bar plot showing the proportion of different subtypes of CD8+ T cells. (I) GSEA of upregulated genes for effector and memory CD8+ T cells (C1) in IL-21+IR group versus IR group. (J) Violin plots showing the expression levels of Gzma, Gzmb, Pdcd1, and Havcr2 of effector and memory CD8+ T cells of IR and IL-21+IR group from scRNA-Seq data. (K) Tumor growth of B16 tumors treated with radiation and/or IL-21 with or without anti–PD-1. Data shown as mean ± SEM in E (n = 5 per group) and K (n = 7 per group). Statistical analysis was carried out using 2-way ANOVA with Tukey’s multiple-comparison test (E and K). For panel J, differences between the 2 groups were assessed using the pairwise.wilcox.test() function in clusterProfiler. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

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PerCP/Cyanine5.5 anti-mouse CD366 (Tim-3)

The PerCP/Cyanine5.5 anti-mouse CD366 (Tim-3) Antibody from BioLegend is a Rat Monoclonal antibody to HAVCR2. This antibody recognizes Mouse antigen. The PerCP/Cyanine5.5 anti-mouse CD366 (Tim-3) Antibody has been shown to work in the following applications: Flow Cytometry.

Description

PerCP/Cyanine5.5 anti-mouse CD366 (Tim-3) [B8.2C12];

Product Specs
ItemPerCP/Cyanine5.5 anti-mouse CD366 (Tim-3)
CompanyBioLegend
Price $382.00Supplier Page View Company Product Page
Catalog Number134012
Quantity100 µg
ApplicationsFC - Quality tested
Concentration0.2mg/ml
Conjugate/TagPerCP/Cyanine5.5
ReactivityMouse
IsotypeRat IgG1, kappa
CloneB8.2C12
HostRat
Antibody TypePrimary
ImmunogenmTim-3 protein/Freund adjuvant
NCBI Full Gene Namehepatitis A virus cellular receptor 2
NCBI Gene AliasesTIM-3, Tim3, Timd3
ClonalityMonoclonal
FormatPhosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
TargetCD366

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(1) NR2F6 deletion revives CAR-T cell function and induces antigen-agnostic immune memory in solid tumorsNature CommunicationsFebruary 27, 2026Dominik Humer, Victoria Klepsch, Dietmar Rieder, Isabel Hölzl, Daniel Schreiber, Viktor Lang, Jiří Koutník, Sebastian Peer, Tajana Sajinovic, Viana Wille, et al.134608), PD-1 bv510 (Biolegend, clone: 29 F.1A12. 135241), Tim3 PerCp-Cy5.5 (Biolegend, clone: B8.2C12, 134011), Lag3 PE-Cy7 (Biolegend, clone: C9B7W, 125225), CD39 PE (Biolegend, clone: Duha59, 143804), CD45 APC

(2) IL-21 enhances the cytotoxicity of intratumoral CD8+ T cells, improving radiation efficacyJCI InsightFebruary 23, 2026Xin-yang Li, Xue-qi Xie, Bao-chao Wei, Xiao-zheng Sun, Min-xin Chen, Ru-fei Liu, Qing-xu Tao, Yi-heng Huang, Qian Wang, Shuang-shuang Ma, et al.123132), CD69 (1:200, BioLegend, 104505), CD279 (1:200, BioLegend, 135231), and CD366 (1:200, BioLegend, 134012). For intracellular staining to evaluate the function of T cells in the TME, single-cell suspensions

(3) MAPKAP Kinase 2 Orchestrates Memory T Cell Inflation in Cytomegalovirus InfectionbioRxivJanuary 23, 2026Eleni Panagioti, Xueyang Yu, Yi Wen Kong, Kristina Makakova, Noe B. Mercado, Sean Edward Lawler, Michael B. Yaffe, Charles H. Cookbiosciences, clone BM8.1, catalog 50-4801, 1:200), Tim3 PerCP/Cy5.5 (BioLegend, clone B8.2C12, catalog 134011, 1:100), KLRG1 FITC (BioLegend, clone 2F1/GKRG1, catalog 138410, 1:100), KLRG1 Brilliant Violet 711

(4) ERBB2 signaling drives immune cell evasion and resistance against immunotherapy in small cell lung cancerNat CommunDecember 9, 2025Lydia Meder, Charlotte I. Orschel, Cyrielle L. Bouchez, Rahil Gholamipoorfard, Claudia V. Orschel, David Stahl, Christoph Kreer, Mirjam Koker, Marieke Nill, Ilayda G. Kocak, et al.(PE-Cy7, clone 10 F.9G2, Biolegend, Cat. # 124313), TIM-3 (PerCP-Cy5.5, clone B8.2C12, Biolegend, Cat. # 134011), Rat IgG2aΚ (Biolegend FITC, Cat. # 400505, PE, Cat. # 400507, PerCP-Cy5.5, Cat. # 400531, APC, Cat.

(5) Tumor-infiltrating nociceptor neurons promote immunosuppressionScience SignalingAugust 5, 2025Anthony C. Restaino, Maryam Ahmadi, Tuany Eichwald, Amin Reza Nikpoor, Austin Walz, Mohammad Balood, Sebastien Talbot, Paola D. Vermeeranti-PD-1-PE-Cy7 (BioLegend, 109110), anti-LAG3–PE (BioLegend, 125208), and anti-TIM3-PerCP/Cyanine5.5 (BioLegend, 134012). These stains were used to identify the CD8+ T cells and to evaluate their expression of various immune

(6) Role of galectin-9 in the development of gestational diabetes mellitusScientific reportsMay 30, 2025Albuayjan HHH, Watanabe M, Sugawara R, Katsuyama E, Mise K, Oi Y, Kanno A, Yang B, Tahara T, Nojima I, et al.(BioLegend Cat# 142607, RRID: AB_2749930), PerCP/Cyanine5.5 anti-mouse CD366 (Tim-3) (BioLegend Cat# 134011, RRID: AB_2632735), and incubated at RT in dark for 15–20 min. The cells were washed by adding 500 µL

(7) Short-term starvation inhibits CD36 N-glycosylation and downregulates USP7 UFMylation to alleviate RBPJ-maintained T cell exhaustion in liver cancerTheranosticsApril 28, 2025Banglun Pan, Siyan Chen, Hao Wu, Xiaoxia Zhang, Zhu Zhang, Dongjie Ye, Yuxin Yao, Yue Luo, Xinyu Zhang, Xiaoqian Wang, et al.(12-9985-82, Invitrogen), anti-mouse-PD1-V450 (75-9981, Cytek, CA, USA), anti-mouse-TIM3-PerCP-Cy5.5 (134011, Biolegend) antibodies. Viability dye used in this study: Fixable Viability Stain 575V (565694, BD Biosciences),

(8) Impact of tumor localization on antitumor immunity with conditionally activated CTLA-4 blockadeJ Immunother CancerApril 2, 2025Marcel Arias-Badia, Chien-Chun Steven Pai, Yee May Lwin, PeiXi Chen, Aahir Srinath, Miho Tanaka, Emily Musser, Andrew Goodearl, Jacob V Gorman, Wendy Ritacco, et al.ASMTNMELM,PE,MBL,TB-5113–1,N/A PRF1,APC,BioLegend,154303,S16009A Ki-67,BV480,BD,566109,B56 FoxP3,PE-Cy7,LT,25-5773-80,FJK-16s TCF-1,A488,BD,567018,S33-966 TIM3,PerCP-Cy5.5,BioLegend,134012,B8.2C12 LAG-3,BV785,BioLegend,125219,C9B7W CD44,BV650,BioLegend,103049,IM7 CD62L,BV570,BioLegend,104433,MEL-14 TNF-α,PE,BioLegend,506305,MP6-XT22 Antibodies

(9) NFAT single-deficient murine T cells reduce the risk of aGvHD while controlling cytomegalovirus infectioniScienceFebruary 21, 2025Nadine Hundhausen, Snigdha Majumder, Yin Xiao, Sigrun S. Haeusl, Helen Goehler, Rishav Seal, Cristina M. Chiarolla, Andreas Rosenwald, Matthias Eyrich, Luka Cicin-Sain, et al.RRID:AB_2564492 anti-mouse KLRG1 (2F1/KLRG1),BioLegend,Cat#: 138421; RRID:AB_2563800 anti-mouse Tim3 (B8.2),BioLegend,Cat#: 134011; RRID:AB_2632735 anti-mouse Lag3 (C9B7W),BioLegend,Cat#: 125210; RRID:AB_10639727 anti-mouse PD-1

(10) Proton radiation boosts the efficacy of mesothelin-targeting chimeric antigen receptor T cell therapy in pancreatic cancerProceedings of the National Academy of Sciences of the United States of AmericaJuly 30, 2024Amit U, Uslu U, Verginadis II, Kim MM, Motlagh SAO, Diffenderfer ES, Assenmacher CA, Bicher S, Atoche SJ, Ben-Josef E, et al.Brilliant Violet 421™ anti-mouse CD279 (PD-1; CAT: 135218; BioLegend), PerCP/Cyanine5.5 anti-mouse CD366 (TIM-3; CAT: 134012; BioLegend), and PE/Dazzle™ 594 anti-mouse CD223 (LAG-3; CAT: 125224; BioLegend). TruStain FcX anti-mouse CD16/32

(11) Tumor Burden Dictates the Neoantigen Features Required to Generate an Effective Cancer VaccineCancer immunology researchApril 2, 2024Garzia I, Nocchi L, Avalle L, Troise F, Leoni G, Seclì L, Antonucci L, Cotugno G, Allocca S, Romano G, et al.BioLegend; catalogue number 329920); TIM3 PerCP-Cyanine 5.5 (clone B8.2C12, BioLegend; catalogue number 134012); Granzyme B FITC (clone GB11, BioLegend; catalogue number 515403); TCF1 APC (clone C63D9, Cell Signaling Technology

(12) Vaccination provides superior in vivo recall capacity of SARS-CoV-2-specific memory CD8 T cellsCell reportsApril 25, 2023Kavazović I, Dimitropoulos C, Gašparini D, Rončević Filipović M, Barković I, Koster J, Lemmermann NA, Babić M, Cekinović Grbeša Đ, Wensveen FM.

(13) Immunization with short peptide particles reveals a functional CD8⁺ T-cell neoepitope in a murine renal carcinoma modelJournal for immunotherapy of cancerDecember 1, 2021He X, Zhou S, Dolan M, Shi Y, Wang J, Quinn B, Jahagirdar D, Huang WC, Tsuji M, Pili R, et al.

(14) Gut Microbiota Modulate CD8 T Cell Responses to Influence Colitis-Associated TumorigenesisCell reportsApril 7, 2020Yu AI, Zhao L, Eaton KA, Ho S, Chen J, Poe S, Becker J, Gonzalez A, McKinstry D, Hasso M, et al.APC-Cy7 clone MEL-14,BioLegend,Cat#104427; AB_S3079S Tim-3 PerCP-Cy5.5 clone BS.2C12,BioLegend,Cat#134011; AB_2632735 PD-1 PE-Cy7 clone RMP1-30,BioLegend,Cat#109109; AB_572016 Lag-3 PE clone C9B7W,BioLegend,Cat#125207;

(15) Enhanced Therapeutic Efficacy of a Novel Oncolytic Herpes Simplex Virus Type 2 Encoding an Antibody Against Programmed Cell Death 1Molecular therapy oncolyticsDecember 20, 2019Zhu Y, Hu X, Feng L, Yang Z, Zhou L, Duan X, Cheng S, Zhang W, Liu B, Zhang K.BioLegend), TIGIT (PE/Cy7, 142107; BioLegend), CD152 (PerCP/Cy5.5, 106315; BioLegend), CD366 (PerCP/Cy5.5, 134011; BioLegend), or CD223 (PerCP/Cy5.5, 125211; BioLegend), or intracellularly stained with IL-2 (APC, 503809;

(16) Flt3L-mobilized dendritic cells bearing H2-Kbm1 apoptotic cells do not induce cross-tolerance to CD8+ T cells across a class I MHC mismatched barrierTransplant international : official journal of the European Society for Organ TransplantationMay 1, 2011Maria-Luisa del Rio, Javier Cote-Sierra, Jose-Ignacio Rodriguez-Barbosa

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Proceedings of the National Academy of Sciences of the United States of America

J Immunother Cancer

J Immunother Cancer

Scientific reports

JCI Insight

PerCP/Cyanine5.5 anti-mouse CD366 (Tim-3) from BioLegend

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BioLegend

Citations (16)

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PerCP/Cyanine5.5 anti-mouse CD366 (Tim-3) from BioLegend

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