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Anti mouse cd45 clone 30 f11

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Anti-mouse CD45 (clone 30-F11) is a lab equipment product used for the detection and analysis of mouse CD45 protein. It is a monoclonal antibody that specifically binds to the CD45 antigen expressed on the surface of mouse cells.

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Lab products found in correlation

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Spelling variants of this product

Anti-CD45 (228 citations) CD45 (30-F11, (72 citations) CD45 (clone 30-F11, (62 citations) Clone 30-F11 (48 citations) Anti-mouse CD45 (39 citations) Anti-CD45 (30-F11, (33 citations) Anti-CD45 (clone 30-F11, (27 citations) Mouse CD45 (clone 30-F11, (5 citations) Anti-mouse CD45 (30-F11, (4 citations) Anti-mouse CD45 antibody clone: 30-F11 (2 citations)

9 protocols using anti mouse cd45 clone 30 f11

1

Multiparametric Flow Cytometry Analysis

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The following antibodies were used for flow cytometry analysis (dilutions are indicated): anti-mouse CD45 (clone 30-F11, 1:100), anti-mouse/human CD11b (clone M1/70, 1:300), anti-mouse Ly6C (clone HK1.4, 1:300), anti-mouse MHCII (clone M5/114.15.2, 1:200), anti-mouse CD64 (clone X54-5/7.1, 1:50), anti-mouse Ly6G (clone 1A8, 1:100), and anti-mouse Tim4 (clone RMT4-54, 1:50), which were purchased from BioLegend, San Diego, CA, USA. Anti-mouse F4/80 (clone A3-1, 1:50) was purchased from BIORAD. The staining for ROS was performed with 0.1 mM of 2,7-dichlorodihydrofluoresceindiacetate (Molecular Probes Invitrogen). Staining for apoptosis and necrosis markers with Annexin V and propidium iodide was performed with MEBCYTO-Apoptosis Kit (MBL International Corporation). Cells were analyzed with BD FACSCanto™ II (BD Bioscience). Flow cytometry analysis was performed using FlowJo software (TreeStar, Ashland, OR, USA).
Graubardt N., Vugman M., Mouhadeb O., Caliari G., Pasmanik-Chor M., Reuveni D., Zigmond E., Brazowski E., David E., Chappell-Maor L., Jung S, & Varol C. (2017). Ly6Chi Monocytes and Their Macrophage Descendants Regulate Neutrophil Function and Clearance in Acetaminophen-Induced Liver Injury. Frontiers in Immunology, 8, 626.
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2

Tumor Immune Profiling by Flow Cytometry

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Tumor lysates were prepared by mincing the tumor in DMEM (Sigma) and incubating in collagenase (Roche) at 37°C for 1 h. After washing in PBS, the cells were filtered through 70‐μm filters (BD Biosciences). 5 × 10 cells were re‐suspended in HBSS (Hank's balanced salt solution, Lonza) supplemented with 0.5% BSA (Sigma). Staining was performed at 4°C for 20 min, with the following antibodies: anti‐mouse CD45 (clone 30‐F11, Biolegend San Diego, CA, USA), anti‐mouse F4/80 (clone BM8, Biolegend San Diego, CA, USA), anti‐mouse cd11c (clone N418, Biolegend San Diego, CA), anti‐mouse I‐A/I‐E (clone M5/114.15.2, Biolegend San Diego, CA, USA); anti‐mouse TNFα (clone MP6‐XT22, BD Pharmingen), anti‐mouse Ly6C (clone HK1.4, eBioscience; San Diego, CA, USA); anti‐mouse NOS2 (clone 6/iNOS/NOS Type II, BD biosciences, San Diego, CA, USA) and mouse IgG2a K (BD biosciences, San Diego, CA, USA). For intracellular staining, Cytofix/Cytoperm and Permwash staining kit (BD Pharmingen) were used. Cells were detected using the BD FACS Canto II cytofluorimeter and analyzed with FlowJo software.
Comunanza V., Corà D., Orso F., Consonni F.M., Middonti E., Di Nicolantonio F., Buzdin A., Sica A., Medico E., Sangiolo D., Taverna D, & Bussolino F. (2016). VEGF blockade enhances the antitumor effect of BRAFV 600E inhibition. EMBO Molecular Medicine, 9(2), 219-237.
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3

Phenotyping Liver Non-Parenchymal Cells

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Non-parenchymal liver cells were incubated with monoclonal antibody 2·4G2 for FcR blocking (BioLegend, San Diego, CA, USA) and then exposed at 4°C to a mixture of the following antibodies (dilutions are indicated):anti-mouse CD45 (clone 30-F11, 1:100), anti-mouse/human CD11b (clone M1/70, 1:300), anti-mouse Ly6C (clone HK1.4, 1:300), anti-mouse MHCII (clone M5/114.15.2, 1:200), anti-mouse CD11c (clone N418, 1:100), anti-mouse CD3ϵ (clone 145-2c11, 1:100), anti-mouse CD8a (clone 53-6.7, 1:100), anti-mouse CD4 (clone GK1.5, 1:100), anti-mouse TCRβ (clone 457-597, 1:100) all were purchased from BioLegend, San Diego, CA. Anti-mouse F4/80 (clone REA 126, 1:100) and anti-mouse Tim4 (clone REA999, 1:100) were purchased from Miltenyi Biotech.
For IL-17A staining, cells were first stained for surface markers, then the cells were fixed and permeabilized prior to intra-cellular staining with IL-17 (Clone TC11-18H10.1, 1:50, BioLegend). Cells were analyzed with BD FACS Canto™ II (BD Bioscience) or sorted with a FACSAria flow cytometer (BD Bioscience). Flow cytometry analysis was performed using FlowJo software (TreeStar, Ashland, OR).
Reuveni D., Brezis M.R., Brazowski E., Vinestock P., Leung P.S., Thakker P., Gershwin M.E, & Zigmond E. (2021). Interleukin 23 Produced by Hepatic Monocyte-Derived Macrophages Is Essential for the Development of Murine Primary Biliary Cholangitis. Frontiers in Immunology, 12, 718841.
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4

Detailed Immune Cell Profiling

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Phenotype analysis was performed with staining performed at 4 °C for 20 min with the following antibodies: anti‐mouse CD45 (clone 30‐F11; Biolegend, San Diego, CA, USA), anti‐mouse F4/80 (clone BM8; Biolegend), anti‐mouse CD11c (clone N418; Biolegend), anti‐mouse Ly6C (clone HK1.4; Thermo Fisher Scientific); anti‐CD11b (clone M1/70; Biolegend, San Diego, CA, USA), anti‐CD206 (clone C068C2; Biolegend), anti‐Ly6G (clone 1A8; Biolegend), anti‐NK1.1 (clone PK136; Biolegend), anti‐CD314 (clone C7; Biolegend), anti‐CD4 (clone GK1.5; BD Pharmigen, San Jose, CA, USA), anti CD8 (clone 53‐6.7; BD Pharmigen); antiPD‐1 (clone 29F.1A12; Biolegend), anti‐PD‐L1 (clone 10F.9G2; Biolegend). Cells were detected using the Cyan ADP flow cytometer (Beckman Coulter, Brea, CA, USA) and data were analyzed with the Summit 4.3 software (Beckman Coulter). Quadrants were set based on isotype control antibody, and cells were gated among total DAPI− cells.
Comunanza V., Gigliotti C., Lamba S., Doronzo G., Vallariello E., Martin V., Isella C., Medico E., Bardelli A., Sangiolo D., Di Nicolantonio F, & Bussolino F. (2023). Dual VEGFA/BRAF targeting boosts PD‐1 blockade in melanoma through GM‐CSF‐mediated infiltration of M1 macrophages. Molecular Oncology, 17(8), 1474-1491.
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5

Comprehensive Immune Cell Profiling

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The following anti-mouse antibodies were purchased from ThermoFisher and used according to manufacturer’s specifications: CD8α (clone 53–6.7, Cat#47–0081-30 or clone Tonbo 2.43, Cat# 50–1886-U100), CD4 (clone RM4–5, Cat #48–0042-82), H-2Kb (clone AF6–88.5.5.3, Cat #12–5958-82), H-2Db (clone 28–14-8, Cat #11–5999-82), anti-mouse isotype IgG2a (clone eBM2a, Cat# 114724–81). Anti-mouse CD45 (clone 30-F11, Cat# 103129) and NKp46 (clone 29A1.4, Cat# 137617) were purchased from Biolegend. Mouse spleens were passed through a 100 μm cell strainer (Corning Life Sciences) and incubated in Red Blood Cell Lysing Buffer Hybri-Max (Sigma-Aldrich) for 3 min at room temperature. For detection of MHC-I, MOE/E6E7 cells were lifted with 1× citric saline to prevent cleavage of MHC-I on cell surface. The prepared cells were incubated with the corresponding panel of antibodies conjugated with unique fluorophores for 30 min to 1 h at room temperature and washed with PBS. Samples were passed through a 35 μm cell strainer (Corning Life Sciences) immediately before analysis on an LSRII flow cytometer (Becton Dickinson) using FACSDiva collection software.
Westrich J.A., Vermeer D.W., Silva A., Bonney S., Berger J.N., Cicchini L., Greer R.O., Song J.I., Raben D., Slansky J.E., Lee J.H., Spanos W.C, & Pyeon D. (2019). CXCL14 suppresses human papillomavirus-associated head and neck cancer through antigen specific CD8+ T cell responses by upregulating MHC-I expression. Oncogene, 38(46), 7166-7180.
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6

Comprehensive Immune Cell Profiling

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The following anti-mouse antibodies were purchased from ThermoFisher and used according to manufacturer’s specifications: CD8α (clone 53–6.7, Cat#47–0081-30 or clone Tonbo 2.43, Cat# 50–1886-U100), CD4 (clone RM4–5, Cat #48–0042-82), H-2Kb (clone AF6–88.5.5.3, Cat #12–5958-82), H-2Db (clone 28–14-8, Cat #11–5999-82), anti-mouse isotype IgG2a (clone eBM2a, Cat# 114724–81). Anti-mouse CD45 (clone 30-F11, Cat# 103129) and NKp46 (clone 29A1.4, Cat# 137617) were purchased from Biolegend. Mouse spleens were passed through a 100 μm cell strainer (Corning Life Sciences) and incubated in Red Blood Cell Lysing Buffer Hybri-Max (Sigma-Aldrich) for 3 min at room temperature. For detection of MHC-I, MOE/E6E7 cells were lifted with 1× citric saline to prevent cleavage of MHC-I on cell surface. The prepared cells were incubated with the corresponding panel of antibodies conjugated with unique fluorophores for 30 min to 1 h at room temperature and washed with PBS. Samples were passed through a 35 μm cell strainer (Corning Life Sciences) immediately before analysis on an LSRII flow cytometer (Becton Dickinson) using FACSDiva collection software.
Westrich J.A., Vermeer D.W., Silva A., Bonney S., Berger J.N., Cicchini L., Greer R.O., Song J.I., Raben D., Slansky J.E., Lee J.H., Spanos W.C, & Pyeon D. (2019). CXCL14 suppresses human papillomavirus-associated head and neck cancer through antigen specific CD8+ T cell responses by upregulating MHC-I expression. Oncogene, 38(46), 7166-7180.
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7

Mouse Tumor Single-Cell Immune Profiling

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Preparation of single-cell suspension of mouse tumor tissues by mechanical grinding. All single-cell suspensions were incubated with rat anti-mouse CD16/CD32 blocking antibody (4 μg/ml) for 15 min after thorough filtration and precipitation, stained with fluorescein-conjugated antibodies, multiple washed with PBS, and resuspended in 7-AAD (exclude non-viable cells). For anti-mouse CD4 (Clone RM4-5, BD Biosciences), anti-mouse CD8 (clone 53-6.7, eBioscience), anti-mouse CD45 (clone 30-F11, Biolegend), anti-mouse CD25 (PC61.5, eBioscience), anti-mouse PD-L1 (clone 10 F.9G2, BioLegend) and anti-mouse CD31 (clone MEC 13.3, BD Biosciences) staining, after incubation for 1 h, cells were washed with PBS for three times (1500 rpm, 5 min each), then detected by flow cytometry (BD FACSCanto II). For FoxP3 staining, after incubation, cells were washed and fixed with 1 ml of fixation & permeabilization solution (BD Biosciences) for 30–60 min, and washed twice with Perm Wash (BD Biosciences). Intracellular staining with anti-FoxP3 antibody (clone FJK-16s, eBioscience) was performed for 1 h. The next steps are the same as described above.
Liu S., Qin T., Liu Z., Wang J., Jia Y., Feng Y., Gao Y, & Li K. (2020). anlotinib alters tumor immune microenvironment by downregulating PD-L1 expression on vascular endothelial cells. Cell Death & Disease, 11(5), 309.
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8

Comprehensive Immune Cell Profiling

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Anti-human CD47 (clone B6H12, BD Biosciences), anti-mouse F4/80 (clone BM8, BioLegend), anti-Sirpα (clone P84, BioLegend), anti-mouse/human CD11b (clone M1/70, BioLegend), anti-mouse CD45 (clone 30-F11, BioLegend), anti-mouse MHC II (clone M5/114.15.2, BioLegend), anti-mouse CD206 (clone C068C2, BioLegend), anti-mouse CD80 (clone 16-10A1, BioLegend), anti-mouse CD86 (clone GL-1, BioLegend), anti-mouse PD-L1 (clone 10F.9G2, BioLegend), anti-mouse Gr1(clone RB6-8C5,BioLegend), anti-human CD14 (clone HCD14, BioLegend), and anti-human CD71 (clone CY1G4, BioLegend; clone OKT9, ThermoFisher; clone L01.1 and clone M-A712, BD Biosciences) were used for FACS analyses. Antibodies were Phycoerythrin (PE)-, PE/Cyanine7, APC, APC/Cyanine7, PerCP/Cyanine5.5, PE/Dazzle™ 594, Alexa Flour® 700 or BV605 conjugated, or fluorophore-conjugated secondary antibodies were used. Annexin V (BD Biosciences), Sytox blue (ThermoFisher), 7-Aminoactinomycin D (7-AAD, ThermoFisher), or Zombie Violet™ Fixable Viability Kit (Biolegend) was used to exclude dead cells. Flow cytometry was performed using the BD LSRFortessa cell analyzers (BD).
Chen J., Cao X., Li B., Zhao Z., Chen S., Lai S.W., Muend S.A., Nossa G.K., Wang L., Guo W., Ye J., Lee P.P, & Feng M. (2021). Warburg Effect Is a Cancer Immune Evasion Mechanism Against Macrophage Immunosurveillance. Frontiers in Immunology, 11, 621757.
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9

Murine Immune Cell Profiling

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Murine samples of spleen tissues, lymph nodes and tumors were collected and prepared as previously described (23) . Cells were stained with the following antibodies: antimouse CD3 (clone 17A2, BioLegend, Cat#100221, RRID:AB_2057374), antimouse CD4 (clone GK1.5, Miltenyi Biotec, Cat#130-121-131, RRID:AB_2752219), antimouse CD8a (clone 53-6.7, BioLegend, Cat#100712, RRID:AB_312751), antimouse CD45 (clone 30-F11, BioLegend, Cat#103108, RRID:AB_312973, BD Biosciences, Cat#557659, RRID:AB_396774), antimouse F4/80 (clone BM8, BioLegend, Cat#123115, RRID:AB_893493), antimouse CD11b (clone M1/70, Miltenyi Biotec, Cat# 130-097-585, RRID:AB_2660136), antimouse Gr-1 (clone RB6-8C5, TONBO, San Diego, CA), antimouse Ly6C (clone HK1.4, BioLegend, Cat#128006, RRID:AB_1186135). Data was acquired using MACS Quant flow cytometer (Miltenyl Biotec, Bergisch Galdbach, Germany) and analyzed using FlowJo software (FlowJo, RRID:SCR_008520).
Mise Y., Hamanishi J., Daikoku T., Takamatsu S., Miyamoto T., Taki M., Yamanoi K., Yamaguchi K., Ukita M., Horikawa N., Abiko K., Murakami R., Furutake Y., Hosoe Y., Terakawa J., Kagabu M., Sugai T., Osakabe M., Fujiwara H., Matsumura N., Mandai M, & Baba T. (2022). Immunosuppressive tumor microenvironment in uterine serous carcinoma via CCL7 signal with myeloid-derived suppressor cells. Carcinogenesis, 43(7).
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