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Cd11b bv510

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CD11b-BV510 is a fluorescently-labeled antibody that binds to the CD11b cell surface marker. CD11b is a membrane-bound integrin subunit that is expressed on the surface of various immune cells, including monocytes, macrophages, and neutrophils. The BV510 fluorescent dye is used to label the CD11b antibody, allowing for the detection and analysis of CD11b-positive cells in flow cytometry applications.

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Lab products found in correlation

Gr1 apc cy7, by BioLegend (5 mentions) Cd45 bv421, by BioLegend (5 mentions) Gallios flow cytometer, by Beckman Coulter (5 mentions) Hyaluronidase, by Merck Group (4 mentions) Cd206 pe cy7, by BioLegend (3 mentions) Cd3 apc cy7, by BioLegend (3 mentions) Ly6g pe cy7, by BioLegend (3 mentions) Nk1.1 apc cy7, by BioLegend (3 mentions) Cd4 pe, by BioLegend (3 mentions) F4 80 fitc, by BioLegend (3 mentions) Murine trustain fcx, by BioLegend (2 mentions) Alexafluor700 f4 80, by Bio-Rad (2 mentions) Percp cy5.5 cd11c, by BD (2 mentions) Siglecf pe cf594, by BD (2 mentions) Cd54 apc, by BioLegend (2 mentions) Pb cd40, by BioLegend (2 mentions) Bv711 cd64, by BioLegend (2 mentions) Bv605 ia ie, by BD (2 mentions) Buv737 cd43, by BD (2 mentions) Cd71 pe, by BioLegend (2 mentions)

Close spelling variants of this product

CD11b (457 citations) Anti-CD11b BV510 (7 citations) BV510-conjugated anti-mouse CD11b (2 citations) CD11b BV510 (M1/70, (2 citations)

19 protocols using cd11b bv510

1

Phenotypic analysis of immune cells

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Tissue was homogenized in serum-free DMEM with 2 mg/ml collagenase type I C (VWR 234153-100MG) and incubated for 1 hour at 37°C. Following incubation, the sample was strained through a 45-μm filter and spun down for 5 minutes at 700 g. Cells were then stained with the following antibodies from BioLegend (1:300 in fluorescence-activated cell sorting [FACS] buffer) for 10 minutes: Alexa-488 CD326 (118210), BV-421 F4/80 (123131), BV-605 CD4 (100547), BV-510 cd11b (101245), Alexa-700 Ly6G (127622), APC CD25 (102012), PE/Cy7 cd11c (117317), APC/Cy7 CD45 (103116), and PE CD45RB (103308). Cells were analyzed on a 5-laser LSR II (Becton Dickson). Details of markers used to quantify individual cells types are presented in S2 Fig.
Lyons J., Ghazi P.C., Starchenko A., Tovaglieri A., Baldwin K.R., Poulin E.J., Gierut J.J., Genetti C., Yajnik V., Breault D.T., Lauffenburger D.A, & Haigis K.M. (2018). The colonic epithelium plays an active role in promoting colitis by shaping the tissue cytokine profile. PLoS Biology, 16(3), e2002417.
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2

Monocyte-derived Macrophage Phenotyping

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Bone marrow cells were flushed from femurs and tibias, centrifuged, then exposed to geys buffer for 5 min on ice, incubated with Fc block (Murine TruStain FcX, Biolegend, 1/50 dilution) for 15 min, incubated with antibodies for 20 min at 4 °C, and washed before sorting CD115+/CD11c-/NK- monocytes using an Influx cell sorter (BD) [68 (link)]. Monocytes were cultured in complete RPMI medium (Thermo Fisher Scientific) and exposed to 100 ng/mL CSF1, 50 µM Q-VD-OPh and 50 µM Ac-YVAD-cmk. After five days, macrophages were studied by flow cytometry using AlexaFluor700-F4/80 (#MCA497A700, Bio-Rad), BV510-CD11b (#101245, Biolegend), PerCP/Cy5.5-CD11c (#560584, BD), APC/Cy7-GR1 (#108424, Biolegend), PE-CF594-SiglecF (#562757, BD), PE-CD71 (#113808, Biolegend), PE/Cy7-CD206 (#141720, Biolegend), APC-CD54 (#116120, Biolegend), PB-CD40 (#124626, Biolegend), BV711-CD64 (#139311, Biolegend), BV605-IA-IE (#563413, BD), BUV737-CD43 (#564398, BD). For the sorting of monocytes, the antibodies used were the following: APC-CD11b (#17-0112-83, Thermo Fisher Scientific), PE/Cy7-CD11c (#561022, BD), FITC-NK (#553164, BD), PerCP/Cy5.5-Ly6C (#560525, BD), AlexaFluor700-Ly6G (#561236, BD), biotin-CD115 (#135507, Biolegend), PE-Streptavidin (#554061, BD). The data were analyzed with FlowJo software v. 10.0.00003.
Solier S., Mondini M., Meziani L., Jacquel A., Lacout C., Berghe T.V., Julé Y., Martinou J.C., Pierron G., Rivière J., Deloger M., Dupuy C., Slama-Schwok A., Droin N., Vandenabeele P., Auberger P., Deutsch E., El-Benna J., Dang P.M, & Solary E. (2023). Caspase Inhibition Modulates Monocyte-Derived Macrophage Polarization in Damaged Tissues. International Journal of Molecular Sciences, 24(4), 4151.
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3

Polarization and Flow Cytometry of BMDMs

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BMDMs were cultured in vitro with M-CSF-conditioned medium. After culturing for 7 days, BMDMs were stimulated for 48 hours with PBS and 100 ng/mL LPS plus 20 ng/mL IFN‐γ to polarize them into M0 and M1 macrophages, respectively. After cell maturation, 0.25% trypsin with 0.02% EDTA (Gibco, Grand Island, USA) was added, and 6‐well plates were plated into incubators for 2 minutes during digestion. Then, DMEM with 10% FBS was added to terminate digestion, the cells were placed into a centrifuge at 300 × g for 5 minutes, and the supernatant was discarded. Cells were resuspended in flow cytometry liquid and blocked with 20 μL of rat serum. Next, 40 μL of the primary antibody mix (BV510-CD11b, 1:200, BioLegend, 101245; APC-F4/80, 1:50, eBioscience, 17-4801-82; PE-CD86, 1:200, BioLegend, 105014) was added and the cells were incubated for 15 minutes on ice in the dark. Then, the cells were washed twice with flow cytometry liquid and assessed on a flow cytometry machine (Beckman Coulter, CA, USA).
Ma Y., Li P., Ju C., Zuo X., Li X., Ding T., Liang Z., Zhang J., Li K., Wang X., Zhu Z., Zhang Z., Song Z., Quan H., Hu X, & Wang Z. (2022). Photobiomodulation Attenuates Neurotoxic Polarization of Macrophages by Inhibiting the Notch1-HIF-1α/NF-κB Signalling Pathway in Mice With Spinal Cord Injury. Frontiers in Immunology, 13, 816952.
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4

Interstitial Macrophages Identification

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Right lungs were digested with the Lung Dissociation kit (Miltenyi Biotec), filtered, erythrocytes were removed using ACK and nucleated cells were collected. Cells were washed with ice-cold PBS, incubated with Fc block (Murine TruStain FcX, Biolegend, 1/50 dilution) for 15 min, incubated with the antibodies for 20 min at 4 °C, washed, then fluorescence was measured with a BD LSRFortessa X-20. Antibodies used were the following: AlexaFluor700-F4/80 (#MCA497A700, Bio-Rad), FITC-CD45 (#103108, Biolegend), BV510-CD11b (#101245, Biolegend), PerCP/Cy5.5-CD11c (#560584, BD), APC/Cy7-GR1 (#108424, Biolegend), PE-CF594-SiglecF (#562757, BD), PE-CD71 (#113808, Biolegend), PE/Cy7-CD206 (#141720, Biolegend), APC-CD54 (#116120, Biolegend), PB-CD40 (#124626, Biolegend), BV711-CD64 (#139311, Biolegend), BV605-IA-IE (#563413, BD), BV650-CD24 (#563545, BD), BUV737-CD43 (#564398, BD). The data were analyzed with FlowJo software v. 10.0.00003. Interstitial macrophages were selected according to their larger size (FSC) and granularity (SSC), on CD45 expression to select leukocytes and GR1-positive cells will be excluded to remove neutrophils, they are equally CD11b high, SiglecF negative, IA-IE positive and CD24 negative.
Solier S., Mondini M., Meziani L., Jacquel A., Lacout C., Berghe T.V., Julé Y., Martinou J.C., Pierron G., Rivière J., Deloger M., Dupuy C., Slama-Schwok A., Droin N., Vandenabeele P., Auberger P., Deutsch E., El-Benna J., Dang P.M, & Solary E. (2023). Caspase Inhibition Modulates Monocyte-Derived Macrophage Polarization in Damaged Tissues. International Journal of Molecular Sciences, 24(4), 4151.
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5

Characterizing Cardiac Immune Populations

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Heart tissue was minced and enzymatically digested using collagenase I (450 U ml−1), collagenase XI (60 U ml−1), DNAse and hyaluronidase (60 U ml−1) (Sigma-Aldrich). Single-cell suspensions were stained with CD45-PerCP/Cy5.5 (clone 30-F11, 1:600, 103132, BioLegend), CD64-APC (clone X54–5/7.1, 1:600, 139305, BioLegend), Ly6G-PE/Cy7 (clone 1A8, 1:600, 127617, BioLegend), Ly6C-FITC (clone HK1.4, 1:600, 128006, BioLegend), CD11b-BV510 (clone M1/70, 1:600, 101245, BioLegend) and DAPI (0.1%, F10347, Thermo Fisher Scientific). Blood samples from Ly6GTdtomato mice were stained with Ly6G-PE/Cy7 (clone 1A8, 1:600, 127617, BioLegend), CD115-BV605 (clone AFS98, 1:600, 135517, BioLegend), CD11b-APC (clone M1/70, 1:600, 101212, BioLegend), CD45-BV711 (30-F11, 1:600, 103147, BioLegend), CD3-APC/Cy7 (clone 17A2, 1:200, 100221, BioLegend), CD19-APC/Cy7 (clone 6D5, 1:300, 115529, BioLegend), B220-APC/Cy7 (clone RA3–6B2, 1:300, 103224, BioLegend), Nk1.1-APC/Cy7 (clone PK136, 1:300, 108724, BioLegend) and DAPI. Data were recorded on an LSRII flow cytometer with FACSDiva 6.1 and analyzed with FlowJo 10 software (BD Biosciences). For qRT–PCR measurements, cells were flow sorted on a FACSAria II (BD Biosciences) into 350 μl of lysis buffer (RNeasy Plus Micro Kit, Qiagen).
Grune J., Lewis A.J., Yamazoe M., Hulsmans M., Rohde D., Xiao L., Zhang S., Ott C., Calcagno D.M., Zhou Y., Timm K., Shanmuganathan M., Pulous F.E., Schloss M.J., Foy B.H., Capen D., Vinegoni C., Wojtkiewicz G.R., Iwamoto Y., Grune T., Brown D., Higgins J., Ferreira V.M., Herring N., Channon K.M., Neubauer S., Sosnovik D.E., Milan D.J., Swirski F.K., King K.R., Aguirre A.D., Ellinor P.T, & Nahrendorf M. (2022). Neutrophils incite and macrophages avert electrical storm after myocardial infarction. Nature cardiovascular research, 1(7), 649-664.
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6

Multiparametric Flow Cytometry of Murine Immune Cells

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Murine spleens and lymph nodes were dissected and passed through 70 µm strainers. After pre‐incubation with Zombie™ dye and Fc‐block (BioLegend), cells were stained with antibodies against B220‐FITC, B220‐PE‐Cy7, CD19‐FITC, CD25‐APC, CD69‐PerCP‐Cy5.5, CD95‐PE (all BD Bioscience), CD4‐PE, CD8‐FITC, CD11b‐BV510, CD21‐FITC, CD23‐APC, CD40‐PE, CD80‐PerCP‐Cy5.5, CD86‐PE and MHC‐II‐PacificBlue (all BioLegend). T cells were stained for interferon (IFN)‐γ‐APC (BioLegend) and IL‐17A‐PE (BD Bioscience) after 4‐h incubation with 50 ng/mL PMA, 0.5 μg/mL ionomycin and 1 μL/mL Golgi‐Plug; regulatory T cells were determined by intracellular staining for Foxp3‐PE (BD Bioscience).
Traub J., Traffehn S., Ochs J., Häusser‐Kinzel S., Stephan S., Scannevin R., Brück W., Metz I, & Weber M.S. (2019). Dimethyl fumarate impairs differentiated B cells and fosters central nervous system integrity in treatment of multiple sclerosis. Brain Pathology, 29(5), 640-657.
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7

Multiparametric Immune Cell Profiling

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1 × 106 cells were Fc blocked (Biolegend101320) and stained for 1 hour, on ice, in the dark, as previously described.20 (link) The following antibodies were used, B220−FITC (BD553087), I Ab PerCP−Cy5.5 (Biolegend116416), Ly6C PE or PECy7 (Biolegend128008, 128071), HLA−DR PECy7 (eBioscience25−9956−41/Biolegend307606), CD11b BV510 (BD562950/Biolegend101263), Ly6G BV605 (BD563005/Biolegend127639), CD11c APC (BD550261), CD3 A700 (eBioscience56−0032−82/Biolegend100216), Live Blue Fluorescent reactive dye (LifeTech L34962), and CD45 BV650 (Biolegend103151).
Jankowska−Gan E., Agashe V.V., Lema D.A., Zhou Y., Gonzalez Bosc L., Sullivan J.A., Greenspan D.S, & Burlingham W.J. (2020). Donor HLA−DR Drives the Development of De Novo Autoimmunity Following Lung and Heart Transplantation. Transplantation Direct, 6(10), e607.
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8

Multiparametric Flow Cytometry of Immune Cells

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Heparinized whole blood was used for flow cytometry using the following antibody cocktail: CD45-PerCP (Clone 30-F11; Biolegend 103130), CD3-eFLUO450 (Clone 17A2; eBioscience 48-0032), NK1.1-PE (Clone PK136; BD557391), Ly6G-APC-CY7 (Clone 1A8; BD560600), CD11b PE-CY7 (Clone M1/70; BD552850), Ly6C-APC (Clone 1G7.G10; Miltenyi 130-093-136), CD19-APC-H7 (Clone 1D3, eBioscience 47-0193) and Siglec-F-PE (Clone E50-2440; BD552126). Further, cells from the whole brain were isolated for FACS. Anesthetized mice were perfused with PBS to remove the peripheral blood. Subsequently, brains were dissected, mechanically dissociated and digested with a collagenase mix (including collagenase IX, collagenase I, DNAse and RPMI-HEPES). Immune cells were separated using a Percoll-gradient and stained with CD45 PerCP (Clone 30-F11; Biolegend 103130), CD11c PE-Cy7 (Clone N418, eBioscience 25-0114)), F4/80 (Clone BM8; Biolegend 123116), CD11b BV510 (Clone M1/70; Biolegend 101245), CD3 BV421 (Clone 17A2; eBioscience 48-0032-82), CD19 BV421 (Clone 6D5; Biolegend 115538), Ly6G BV421 (Clone eBio927; eBioscience 48-3172), MHC II-APC-eFluor780 (Clone M5/114.15.2; eBioscience 47-5321-82). All samples were measured with a FACS-Canto II (BD Biosciences). Results were analysed with FACSdiva version 8 (BD Biosciences).
Kerkhofs D., van Hagen B.T., Milanova I.V., Schell K.J., van Essen H., Wijnands E., Goossens P., Blankesteijn W.M., Unger T., Prickaerts J., Biessen E.A., van Oostenbrugge R.J, & Foulquier S. (2020). Pharmacological depletion of microglia and perivascular macrophages prevents Vascular Cognitive Impairment in Ang II-induced hypertension. Theranostics, 10(21), 9512-9527.
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9

Immune Cell Isolation from Surgical Specimens

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Immediately post-excision, surgical specimens were placed in isotonic saline on ice and rapidly processed. Briefly, tissues were weighed, placed onto a piece of 250 μm Nitex mesh and dissociated with a mortar and pestle in HBSS + 10% FBS. Cells were washed 2X with HBSS + 10% FBS and after the final wash were layered over a Ficoll-Paque gradient and centrifuged according to the manufacturer's instructions (GE Healthcare, Uppsala, Sweden). Leukocytes were collected from the interface, washed, and counted. Cells were then incubated with Human FcR Receptor Binding Inhibitor (eBioscience, San Diego, CA) to minimize non-specific antibody binding. Cells were then stained with directly-conjugated antibodies for multi-color flow cytometry analysis, which included anti-human CD66b-FITC (BioLegend, San Diego, CA), CD14-PE, CD16-APC, CD33-PE-Cy5, HLA-DR-PE-Cy7, CD45-eFluor450, CD3-APC-Cy7, CD11b-BV510 and CD11c-BV605. All fluorochrome-conjugated antibodies were purchased from either BD or eBioscience unless otherwise noted. To exclude dead cells from analysis, a Live/Dead Fixable Stain Kit (Life Technologies, Eugene, OR) was used according to the manufacturer's instructions. Controls included cells stained with isotype control antibodies to assess the degree of non-specific staining. Analysis was performed using BD FACSDiva software with cells gated on the live CD45+ leukocyte population.
Heim C.E., Vidlak D., Scherr T.D., Hartman C.W., Garvin K.L, & Kielian T. (2015). Interleukin-12 promotes myeloid-derived suppressor cell (MDSC) recruitment and bacterial persistence during S. aureus orthopedic implant infection. Journal of immunology (Baltimore, Md. : 1950), 194(8), 3861-3872.
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10

Multiparameter Flow Cytometry Panel

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PD-1 FITC (1:300; J43; eBioscience), NK1.1 PE (1:300; PK136; eBioscience), CD3e PE-Dazzle594 (1:300; 145-2C11; BioLegend), CD3e PE (1:200; 145-2C11; eBioscience), FoxP3 PerCP-Cy5.5 (1:200; FJK-16s; eBioscience), CD8a PE-Cy7 (1:2000; 53-6.7; eBioscience), CD8a APC (1:600; 53-6.7; eBioscience), CD8a PerCP-Cy5.5 (1:400; 53-6.7; eBioscience), TCRb AF700 (1:100; H57-597; BioLegend), CD45 BV785 (1:1000; 30-F11; BioLegend), CD4 BV711 (1:400; GK1.5; BioLegend), CD19 BV650 (1:400; 6D5; BioLegend), CD19 PE (1:500; 1D3; eBioscience), CD11b BV605 (1:1000; M1/70; BioLegend), CD11b BV510 (1:400; M1/70; BioLegend), CD11c BV605 (1:400; N418; BioLegend), Siglec-F PE (1:300; E50-2440; BD Biosciences), F4/80 APC (1:200; BM8; BioLegend), Ly-6G AF 700 (1:300; 1A8; BioLegend), MHC II BV650 (1:4000; M5/114.15.2; BioLegend), CD64 BV421 (1:200; X54-5/7.1; BioLegend), TNF FITC (1:300; MP6-XT22; BioLegend), IFNg PE-Cy7 (1: 1000; XMG1.2; BD Biosciences), Eomes PE (1:200; W17001A; BioLegend), Ki-67 BV 650 (1:200, 11F6; Biolegend), GzmB FITC (1:100; GB11; BioLegend). Gating strategy is depicted in Supplementary Fig. 6d.
Frey N., Tortola L., Egli D., Janjuha S., Rothgangl T., Marquart K.F., Ampenberger F., Kopf M, & Schwank G. (2022). Loss of Rnf31 and Vps4b sensitizes pancreatic cancer to T cell-mediated killing. Nature Communications, 13, 1804.
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