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Cd11b icrf44

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CD11b (ICRF44) is an integrin alpha M chain that is expressed on the surface of various immune cells, including monocytes, macrophages, and neutrophils. It plays a core role in cell adhesion, migration, and phagocytosis.

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Lab products found in correlation

Cd14 hcd14, by BioLegend (3 mentions) Cd163 ghi 61, by BioLegend (2 mentions) Hla dr l243, by BioLegend (2 mentions) Cytofix cytoperm fixation permeabilization kit, by BD (2 mentions) Live dead fixable aqua dead cell stain, by Thermo Fisher Scientific (2 mentions) Rpa t8, by BioLegend (2 mentions) Hcd56, by BioLegend (2 mentions) Rpa t4, by BioLegend (2 mentions) Streptavidin apc cy7, by BioLegend (2 mentions) Clone rpa 2, by BioLegend (2 mentions) Cd7 124 1d1, by Thermo Fisher Scientific (2 mentions) Cd10 sn5c, by Thermo Fisher Scientific (2 mentions) Cd20 2h7, by Thermo Fisher Scientific (2 mentions) Anti human cd34 apc 8g12, by BD (2 mentions) Anti cd38 pe cy7 hit2, by BioLegend (2 mentions) Anti cd90 fitc 5e10, by BD (2 mentions) Anti cd45ra pb mem 56, by Thermo Fisher Scientific (2 mentions) Anti cd123 pe 7g3, by BD (2 mentions) Anti human ki 67 bv605 antibody, by BioLegend (2 mentions) Cd19 hib19, by BioLegend (2 mentions)

6 protocols using cd11b icrf44

1

Phenotypic Profiling of Monocytes and DCs

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1x106 cryopreserved PBMC were thawed then washed twice in FACS buffer and surface stained using the following antibody cocktail: CD14 (M5E2, Biolegend), CD16 (3G8, Biolegend), CD11b (ICRF44, Biolegend), HLA-DR (L243, Biolegend), CD40 (5C3, BD Pharmingen), CD62L (DREG-56, BD Pharmingen), CD64 (10.1, Biolegend), CD163 (GHI/61, Biolegend), CXCR6 (K041E5, Biolegend), IFNAR (85228, R&D Systems), CD11c (3.9, ThermoFisher Scientific) and CD123 (6H6, Biolegend) for the detection of various monocytic and dendritic cell subsets. Samples were then acquired on the Attune NxT acoustic focusing cytometer (Life Technologies). Data were analyzed using FlowJo v10 (TreeStar, Ashland, OR USA).
Zulu M.Z., Sureshchandra S., Pinski A.N., Doratt B., Shen W, & Messaoudi I. (2021). Obesity Correlates With Pronounced Aberrant Innate Immune Responses in Hospitalized Aged COVID-19 Patients. Frontiers in Immunology, 12, 760288.
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2

Flow Cytometric Immunophenotyping of Hematopoietic Progenitor Cells

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Cells were thawed at 37 °C, washed with sterile PBS and incubated with biotinylated anti-human lineage antibodies directed against CD2 (clone RPA-2.10, BioLegend), CD3 (HIT3a, 300304, BioLegend), CD4 (RPA-T4, BioLegend), CD7 (124-1D1, eBioscience), CD8a (RPA-T8, BioLegend), CD10 (SN5c, eBioscience), CD11b (ICRF44, BioLegend), CD14 (HCD14, BioLegend), CD19 (HIB19, BioLegend), CD20 (2H7, eBioscience), CD56 (HCD56, BioLegend) and GPA (HIR2, BioLegend) and LIVE/DEAD Fixable Aqua Dead Cell Stain (L-34957, Life Technologies). For gating on hematopoietic progenitor cells, this was followed by secondary staining with anti-human CD34–APC (8G12, BD Biosciences), anti-CD38–PE/Cy7 (HIT2, BioLegend), anti-CD90–FITC (5E10, BD Biosciences), anti-CD45RA-PB (MEM-56, Thermo Fisher Scientific), anti-CD123–PE (7G3, BD Biosciences) and streptavidin-APC/Cy7 (BioLegend). Intracellular staining with anti-human Ki-67–BV605 antibody (BioLegend) was performed using the BD Cytofix/Cytoperm Fixation/Permeabilization kit. Samples were run on an LSR II flow cytometer (BD Biosciences), and recorded events were analyzed with FlowJo 10 software (BD). Individual fluorescence-minus-one controls were used to determine gating. HSCs were identified as lineage−CD34+CD38−CD45RAloCD90+ cells, CMPs as lineage−CD34+CD38intCD45 RA−CD123int cells and GMPs as lineage−CD34+CD38intCD45RA+CD123int cells.
Rohde D., Vandoorne K., Lee I.H., Grune J., Zhang S., McAlpine C.S., Schloss M.J., Nayar R., Courties G., Frodermann V., Wojtkiewicz G., Honold L., Chen Q., Schmidt S., Iwamoto Y., Sun Y., Cremer S., Hoyer F.F., Iborra-Egea O., Muñoz-Guijosa C., Ji F., Zhou B., Adams R.H., Wythe J.D., Hidalgo J., Watanabe H., Jung Y., van der Laan A.M., Piek J.J., Kfoury Y., Désogère P.A., Vinegoni C., Dutta P., Sadreyev R.I., Caravan P., Bayes-Genis A., Libby P., Scadden D.T., Lin C.P., Naxerova K., Swirski F.K, & Nahrendorf M. (2021). Bone marrow endothelial dysfunction promotes myeloid cell expansion in cardiovascular disease. Nature cardiovascular research, 1(1), 28-44.
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3

Flow Cytometric Immunophenotyping of Hematopoietic Progenitor Cells

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Cells were thawed at 37 °C, washed with sterile PBS and incubated with biotinylated anti-human lineage antibodies directed against CD2 (clone RPA-2.10, BioLegend), CD3 (HIT3a, 300304, BioLegend), CD4 (RPA-T4, BioLegend), CD7 (124-1D1, eBioscience), CD8a (RPA-T8, BioLegend), CD10 (SN5c, eBioscience), CD11b (ICRF44, BioLegend), CD14 (HCD14, BioLegend), CD19 (HIB19, BioLegend), CD20 (2H7, eBioscience), CD56 (HCD56, BioLegend) and GPA (HIR2, BioLegend) and LIVE/DEAD Fixable Aqua Dead Cell Stain (L-34957, Life Technologies). For gating on hematopoietic progenitor cells, this was followed by secondary staining with anti-human CD34–APC (8G12, BD Biosciences), anti-CD38–PE/Cy7 (HIT2, BioLegend), anti-CD90–FITC (5E10, BD Biosciences), anti-CD45RA-PB (MEM-56, Thermo Fisher Scientific), anti-CD123–PE (7G3, BD Biosciences) and streptavidin-APC/Cy7 (BioLegend). Intracellular staining with anti-human Ki-67–BV605 antibody (BioLegend) was performed using the BD Cytofix/Cytoperm Fixation/Permeabilization kit. Samples were run on an LSR II flow cytometer (BD Biosciences), and recorded events were analyzed with FlowJo 10 software (BD). Individual fluorescence-minus-one controls were used to determine gating. HSCs were identified as lineage−CD34+CD38−CD45RAloCD90+ cells, CMPs as lineage−CD34+CD38intCD45 RA−CD123int cells and GMPs as lineage−CD34+CD38intCD45RA+CD123int cells.
Rohde D., Vandoorne K., Lee I.H., Grune J., Zhang S., McAlpine C.S., Schloss M.J., Nayar R., Courties G., Frodermann V., Wojtkiewicz G., Honold L., Chen Q., Schmidt S., Iwamoto Y., Sun Y., Cremer S., Hoyer F.F., Iborra-Egea O., Muñoz-Guijosa C., Ji F., Zhou B., Adams R.H., Wythe J.D., Hidalgo J., Watanabe H., Jung Y., van der Laan A.M., Piek J.J., Kfoury Y., Désogère P.A., Vinegoni C., Dutta P., Sadreyev R.I., Caravan P., Bayes-Genis A., Libby P., Scadden D.T., Lin C.P., Naxerova K., Swirski F.K, & Nahrendorf M. (2021). Bone marrow endothelial dysfunction promotes myeloid cell expansion in cardiovascular disease. Nature cardiovascular research, 1(1), 28-44.
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4

Multiparameter Flow Cytometry of Macaque Immune Cells

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The following fluorochrome conjugated monoclonal antibodies reactive with macaque cells were used for flow cytometry studies: CD3 (SP34-2, BD Biosciences [BD]), CD4 (L200, BD), CD8 (SK1, BD), CD95 (DX2, BD), CD28 (CD28.2, BioLegend), CCR5 (3A9, BD), CXCR5 (MU5UBEE, Invitrogen), PD-1 (EH12.2H7, BioLegend), ICOS (C398.4A, BioLegend), CCR7 (150503, BD), α4β7 (A4B7, NHP Reagent Resource), LAG3 (3DS223H, eBioscience), Tim-3 (F38-2E2, BioLegend), TIGIT (MBSA43, Invitrogen), CD20 (2H7, BioLegend), CD19 (J3-119, Beckman Coulter), HLA-DR (L243, BioLegend), CD10 (HI10A, BioLegend), CD21 (B-ly4, BD), CD27 (O323, BioLegend), IgD (IADB6, Southern Biotech), IgG (G18-145, BD), IL-21R (2G1-K12, BioLegend), Ki-67 (B56, BD), BCL6 (IG191E/A8, BioLegend), CD80 (2D10, BioLegend), CD56 (B159, BD), CD45 (D058-1283, BD), CD14 (MoP9, BD), CD16 (3G8, BD), CCR2 (48607, R&D Systems), CD11b (ICRF44, BioLegend), CD11c (3.9, Invitrogen), PD-L1 (29E.2A3, BioLegend), PD-L2 (24F.10C12, BioLegend), CD80 (2D10, BioLegend), CD86 (FUN-1, BD), CD141 (1A4, BD), CD163 (GHI/61, BioLegend), and CD123 (7G3, BD).
Kvistad D., Pallikkuth S., Sirupangi T., Pahwa R., Kizhner A., Petrovas C., Villinger F, & Pahwa S. (2021). IL-21 enhances influenza vaccine responses in aged macaques with suppressed SIV infection. JCI Insight, 6(20), e150888.
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5

Isolation and Sorting of Plasma Cells

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BM aspirates were collected in heparin-containing tubes, and mononuclear cells were isolated with the Ficoll Paque Plus kit (GE Healthcare). Mononuclear cells were enriched for CD138 with magnetic beads (Miltenyi Biotec) and then stained by CD3 (HIT3a), CD11b (ICRF44), and CD138 (MI15) (BioLegend). CD3–CD11b–CD138+ plasma cells were sorted on a FACSAria III (BD).
Isshiki Y., Oshima M., Mimura N., Kayamori K., Miyamoto-Nagai Y., Seki M., Nakajima-Takagi Y., Kanamori T., Iwamoto E., Muto T., Tsukamoto S., Takeda Y., Ohwada C., Misawa S., Ikeda J.I., Sanada M., Kuwabara S., Suzuki Y., Sakaida E., Nakaseko C, & Iwama A. (2022). Unraveling unique features of plasma cell clones in POEMS syndrome with single-cell analysis. JCI Insight, 7(20), e151482.
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6

Cytotoxicity Assay with CellTrace Violet

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CellTrace™ Violet (Invitrogen)-labeled targets were incubated at the indicated ratios with effector cells for 3 hours. Where required, targets were labeled with an antibody mix containing PeCy7-conjugated mouse anti-human CD3 (OKT3, Biolegend), CD56 (5.1H11, Biolegend), CD11b (ICRF44, Biolegend), CD14 (HCD14, Biolegend) and CD16 (B73.1, Biolegend) mAbs to allow discrimination between CD19+ and CD19- mononuclear cells. As controls, targets and effectors alone were simultaneously incubated to determine spontaneous cell death. Where indicated, targets were pre-incubated with αGalCer or vehicle at 37°C for 4 hr before addition of the effector cells. Cells were then harvested and 7-AAD was added prior to flow cytometric analysis on BD Fortessa Flow Cytometer, using BD FACSDiva software version 6.0. Specific cytotoxic activity was determined as ((% sample (7-AAD+, Violet+) − % spontaneous (7-AAD+, Violet+)) / (100 - %spontaneous (7-AAD+, Violet+)) x 100. All assays were run in duplicates or triplicates. Data analysis was performed using FlowJo 10.2.
Rotolo A., Caputo V.S., Holubova M., Baxan N., Dubois O., Chaudhry M.S., Xiao X., Goudevenou K., Pitcher D.S., Petevi K., Kachramanoglou C., Iles S., Naresh K., Maher J, & Karadimitris A. (2018). Enhanced Anti-lymphoma Activity of CAR19-iNKT Cells Underpinned by Dual CD19 and CD1d Targeting. Cancer Cell, 34(4), 596-610.e11.
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