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Anti cd11b percp

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Anti-CD11b-PerCP is a fluorochrome-conjugated antibody that binds to the CD11b antigen. CD11b is a cell surface glycoprotein that is expressed on the surface of various immune cells such as monocytes, macrophages, and neutrophils. The PerCP fluorochrome allows for the detection and quantification of CD11b-positive cells using flow cytometry.

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Lab products found in correlation

Anti cd4 apc, by BioLegend (1 mentions) Anti cd11c pe, by BD (1 mentions) Anti cd69 fitc, by BD (1 mentions) Sytox blue, by Thermo Fisher Scientific (1 mentions) Pacific blue anti cd8, by Thermo Fisher Scientific (1 mentions) Trustain fcx, by BioLegend (1 mentions) Facsaria iiu, by BD (1 mentions) Brilliant violet buffer, by BD (1 mentions) Mouse fc block, by BioLegend (1 mentions) Ultracomp beads, by Thermo Fisher Scientific (1 mentions) Sorp lsr 2, by BD (1 mentions) Anti ly6c fitc, by BD (1 mentions) Cd16 32, by BioLegend (1 mentions) Anti cd45 apc cy7, by BioLegend (1 mentions) Anti cd11c pe, by BioLegend (1 mentions) Anti cd45.2 pe, by BioLegend (1 mentions) Anti f4 80 bv421, by BioLegend (1 mentions) Anti gr 1 apc, by Thermo Fisher Scientific (1 mentions) Collagenase d, by Merck Group (1 mentions) Dnase 1, by Merck Group (1 mentions)

9 protocols using anti cd11b percp

1

Analyzing Immune Cells in BALF

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After sacrifice, BALF was obtained as previously described (5 (link)). The cells were pelleted and analyzed by flow cytometry using: FITC anti-CD69 (cat# 561929; BD Pharmingen), PerCP anti-CD8a (cat# 45-0081-82; eBioscience), APC anti-CD4 (cat# 100516; BioLegend), PE anti-CD11c (cat#553802, BD Pharmingen), anti-Siglec-H (cat# MCA4647GA, AbD Serotec, Raleigh, NC, USA), APC anti-CD317 (cat# 127015, BioLegend), PerCP anti-CD11b (cat# 101229, BioLegend), FITC anti-rat IgG (cat# 712-095-150, Jackson ImmunoResearch Laboratories). In general, 106 cells were blocked with the anti-FcR mAb 2.4G2, stained with the indicated antibodies for 20 min at 4°C and then washed and resuspended in FACS buffer for analysis. Flow cytometry was performed on a BD FACSCalibur™. Data analysis was performed using BD CellQuest™ Pro software. Cell-free BALF was subjected to cytokine analysis by cytometric bead arrays (CBA) or MPO–DNA complex analysis. The lung was snap frozen in OCT compound and examined for in vivo NET formation.
Akk A., Springer L.E, & Pham C.T. (2016). Neutrophil Extracellular Traps Enhance Early Inflammatory Response in Sendai Virus-Induced Asthma Phenotype. Frontiers in Immunology, 7, 325.
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2

Antibody Staining with Fc Receptor Blockade

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Antibody staining was done in presence of Fc receptor blockade (TruStain fcX BioLegend, #101320) in FACS buffer (1× PBS with 1% FBS). SytoxBlue (Thermo Fisher Scientific, S34857) was used for exclusion of dead cells. Antibodies used for flow cytometry were: PerCP-anti-CD11b (Biolegend, #101230), Pacific Blue-anti-CD8 (ebioscience, #48-0081-82). The cells were stained in FACS buffer containing antibodies at 4 °C for 30 min, washed and resuspended in FACS buffer. A FACSAria IIu (BD Biosciences, Heidelberg) and FlowJo software (TreeStar) were used for acquisition and analysis.
Schröder M., Krötschel M., Conrad L., Naumann S.K., Bachran C., Rolfe A., Umansky V., Helming L, & Swee L.K. (2018). Genetic screen in myeloid cells identifies TNF-α autocrine secretion as a factor increasing MDSC suppressive activity via Nos2 up-regulation. Scientific Reports, 8, 13399.
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3

Flow Cytometry Analysis of SVF Cells

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SVF cells were purified as described above. Flow cytometry analyses were performed using the following anti-mouse antibodies: PerCP anti CD11b (Biolegend), FITC antiF4/80 (BioRAD). Single cell suspensions were stained with Aqua Zombie dye, washed, pre-blocked with mouse FcBlock (Biolegend), and stained with antibody cocktails in the presence of Brilliant Violet buffer (BD Biosciences). Ultracomp beads (eBioscience) stained with abovementioned antibodies and ArC beads (Life Technologies) stained with Aqua Zombie dye were used for compensation in FACSDIVA. All data were acquired on a SORP LSRII (BD Biosciences). At least 20,000 events were collected from each sample. Manual data analysis was performed by a blinded investigator using FlowJo 10. Results presented are from three independent experiments, with statistical significance calculate by two-tailed Student's t test.
Cederquist C.T., Lentucci C., Martinez-Calejman C., Hayashi V., Orofino J., Guertin D., Fried S.K., Lee M.J., Cardamone M.D, & Perissi V. (2016). Systemic insulin sensitivity is regulated by GPS2 inhibition of AKT ubiquitination and activation in adipose tissue. Molecular Metabolism, 6(1), 125-137.
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4

Isolation and Analysis of Murine Myeloid Cells

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Peripheral blood from MФ-PP2Acα+/+ mice and MФ-PP2Acα−/− mice was collected with EDTA as anticoagulant. After blocking with CD16/32 (cat: 101330, Biolegend) for 30 min, blood samples were incubated with PE-anti-Ly6g (cat: 83122-60-25, BioGems, Westlake Village, CA, USA), FITC-anti-Ly6c (cat: 553104, BD Biosciences, New Jersey, USA), and PerCP-anti-CD11b (cat: 101230, Biolegend, San Diego, USA) for 20 min. Then the red blood cells were lysed with RCLB. After centrifugation at 450 g for 5 min, cells were detected and analyzed with BD Canto II Flow Cytometer and the FlowJo software.
Liang Y., Sun X., Wang M., Lu Q., Gu M., Zhou L., Hou Q., Tan M., Wang S., Xue X, & Dai C. (2021). PP2Acα promotes macrophage accumulation and activation to exacerbate tubular cell death and kidney fibrosis through activating Rap1 and TNFα production. Cell Death and Differentiation, 28(9), 2728-2744.
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5

Profiling Lung-Infiltrating Immune Cells

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When the lung-infiltrating cells were analyzed, anti-CD45-APC-Cy7 (BioLegend) were intravenously injected (3 μg/mouse) before euthanasia to exclude the blood-circulating immune cells. Cells from lung or spinal cord tissues were prepared by enzymatic digestion with 2.5 mg/ml collagenase D (Sigma-Aldrich) and 0.1 mg/ml DNase I (Sigma-Aldrich) for 30 min at 37°C. For neutrophils, monocytes, macrophages, and CD4+ T cell staining, the cells or tissues were stained with anti-Ly6c-AF488 (BioLegend), anti-CD45.2-PE (BioLegend), anti-CD11c-PE (BioLegend), anti-CD11b-PerCP (BioLegend), anti-Gr-1-APC (eBioscience), anti-CD4-APC (TONBO), and anti-F4/80-BV421 (BioLegend).
Miyashita Y., Kouwaki T., Tsukamoto H., Okamoto M., Nakamura K, & Oshiumi H. (2021). TICAM-1/TRIF associates with Act1 and suppresses IL-17 receptor–mediated inflammatory responses. Life Science Alliance, 5(2), e202101181.
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6

Comprehensive Immune Cell Profiling in Sepsis

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Mice were sacrificed and spleens were harvested at 24h after CLP. Splenocytes were stained with anti-CD3-Alexa 700 (BD), anti-CD4-PB (Biolegend, clone RM4–5), anti-CD8-PO (Invitrogen, clone MCD0830), anti-CD44-PerCP (Biolegend, clone IM7), anti-CD62L-PE (BD), anti-CD28-PE-Cy7 (Biolegend, clone E18) and anti-CD25-APC-Cy7 (BD). For detection of cell apoptosis, splenocytes were stained with a FITC Annexin V apoptosis detection kit with 7-AAD (Biolegend). Anti-Bcl-xL (54H6) and Bcl-2 (Biolegend, clone BCL/10C4) were used to detect engagement of the mitochondrial pathway of apoptosis while anti-CD95 (Biolegend, clone DX2) and anti-TNFR Type Ⅰ (Biolegend, clone 55R-286) were stained to detect expression of death receptors on T cells. Cells were intracellularly stained with anti-Ki-67 (Biolegend, clone 16A8) to assay for cell proliferation. Tregs were identified via intracellular staining for Foxp3-FITC (Ebioscience, clone FJK-16S) using the Foxp3 staining kit (Ebioscience). B cells were stained with anti-CD19-FITC (Biolegend). NK cells were stained with anti-NK1.1-PE (Ebioscience). Dendritic cells were stained with anti-CD11c-PE-Cy7 (BD). Neutrophils were stained with anti-Gr-1-Alexa 700 (Biolegend) and anti-CD11b-PerCP (Biolegend). Accucheck Counting Beads (Thermo Fisher Scientific) were added after staining to calculate the absolute number of cells per spleen.
Sun Y., Xie J., Anyalebechi J.C., Chen C.W., Sun H., Xue M., Liang Z., Morrow K.N., Coopersmith C.M, & Ford M.L. (2020). CD28 agonism improves survival in immunologically experienced septic mice via IL-10 released by Foxp3+ regulatory T cells. Journal of immunology (Baltimore, Md. : 1950), 205(12), 3358-3371.
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7

Isolation and Culture of Mouse BMSCs

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C57BL/6J mice were anesthetized with pentobarbital and then sacrificed. The tibias and femurs of the mice were separated, and the ends of the tibias and femurs were cut. The bone marrow was flushed out with a 10 ml syringe with a 25-gauge needle filled with 10 ml of 1× PBS. The cell suspensions were centrifuged at 700 × g for 5 minutes at 4°C, and the cells were resuspended in 500 μl of 1× PBS. Then, the cell suspensions were incubated with anti-Sca-1-PE (BioLegend, USA), anti-CD29-FITC (BioLegend, USA), anti-CD45-PerCP (BioLegend, USA), and anti-CD11b-PerCP (BioLegend, USA) antibodies for 30 minutes at 4°C. The mouse BMSCs (Sca-1+CD29+CD45-CD11b-) were sorted by fluorescence-activated cell sorting (FACS) with a FACSAria (BD Biosciences, USA) and cultured with α-MEM culture medium (Cellmax, China) with 10% fetal bovine serum (FBS) and 1% streptomycin (Gibco, USA) and penicillin (Gibco, USA) at 37°C in a humidified atmosphere of 5% CO2.
Wang J., Xie X., Li H., Zheng Q., Chen Y., Chen W., Chen Y., He J, & Lu Q. (2024). Vascular endothelial cells-derived exosomes synergize with curcumin to prevent osteoporosis development. iScience, 27(4), 109608.
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8

NK Cell Receptor Profiling Protocol

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Splenocytes were suspended with FACS buffer (1 mM EDTA, 0.01% sodium azide, 0.1% Bovine Serum Albumin (BSA), 0.02 M phosphate, 0.15 M NaCl, pH 7.2) and then stained for 20 min with anti-FcR/anti-CD16 + CD32/Fc Block (eBioscience) and the following fluorophore-conjugated antibodies for NK cell receptor profiling: anti-CD19-APC (BioLegend), anti-CD3-Pac Blue (BioLegend), anti-NK1.1-PE Cy7 (BioLegend), anti-NKG2A-PE (BioLegend), anti-CD11b-PerCP (BioLegend), anti-CD27-AF700 (BioLegend), anti-CX3CR1-BV510 (BioLegend), anti-CD107a-APC Cy7 (BioLegend), and anti-NKG2D-FITC (BioLegend). After staining, cells were washed three times with 200 µl FACS buffer. Data were acquired on a LSRII instrument (BD Biosciences). Analysis was performed using FlowJo software, version 10.0.8.
Menees K.B., Earls R.H., Chung J., Jernigan J., Filipov N.M., Carpenter J.M, & Lee J.K. (2021). Sex- and age‐dependent alterations of splenic immune cell profile and NK cell phenotypes and function in C57BL/6J mice. Immunity & Ageing : I & A, 18, 3.
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9

Tumor Immune Cell Profiling by Flow Cytometry

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Tumor immune cells isolated as described above were resuspended at 106 cells/100 μl in FACS buffer (2% BSA, 2% goat serum in PBS). Cells were immunolabeled with the following fluorochrome-conjugated cell surface antibodies: anti-CD45 PE/Cy5 (0.25 μg/100 μl), anti-CD11b PerCP (0.25 μg/100 μl), anti-CD4 FITC (0.25 μg/100 μl), anti-CD8 PE (0.25 μg/100 μl), anti-CD25 PE/Cy7 (0.5 μg/100 μl), and anti-FOXP3 Alexa Fluor 647 (5 μl/100 μl) all purchased from BioLegend. Single cells were prepared for flow cytometry as previously described [23 ] or according to the manufacturer’s protocol (BioLegend, for FOXP3). Cell-associated fluorescence was acquired and analyzed using the BD LSR II cytometer and TreeStar Inc. FlowJo software, respectively.
Obr A.E., Kumar S., Chang Y.J., Bulatowicz J.J., Barnes B.J., Birge R.B., Lazzarino D.A., Gallagher E., LeRoith D, & Wood T.L. (2018). Insulin-like growth factor receptor signaling in breast tumor epithelium protects cells from endoplasmic reticulum stress and regulates the tumor microenvironment. Breast Cancer Research : BCR, 20, 138.
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