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M1 70

Manufactured by Miltenyi Biotec
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The M1/70 is a lab equipment product from Miltenyi Biotec. It is a monoclonal antibody that recognizes the CD11b antigen, also known as the Mac-1 integrin. This antibody is commonly used for the identification and isolation of myeloid cells, such as macrophages and granulocytes, in various research applications.

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Lab products found in correlation

Lung dissociation kit, by Miltenyi Biotec (2 mentions) Sh800 flow cytometer, by Sony (2 mentions) Cd45 microbeads, by Miltenyi Biotec (2 mentions) Facsverse flow cytometer, by BD (2 mentions) Dcn228, by Miltenyi Biotec (1 mentions) Hi149, by Miltenyi Biotec (1 mentions) Rea968, by Miltenyi Biotec (1 mentions) Hcd14, by BioLegend (1 mentions) Diva software, by BD (1 mentions) Rea641, by Miltenyi Biotec (1 mentions) Rea1162, by Miltenyi Biotec (1 mentions)

Close spelling variants of this product

CD11b-APC (clone M1/70, APC)-conjugated CD11b mouse clone M1/70.15.11.5

8 protocols using m1 70

1

Isolation and Characterization of Lung Macrophages

Cited 1 time
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To validate the performance of CPM we sorted the population of macrophages from the lungs of various CC mice (Supplementary Table 1). To address this, the lungs were dissociated into single cell suspensions using Miltenyi Biotec lung dissociation kit (130-095-927), according to manufacturer’s instructions. Isolated lung cells were then enriched for CD45+ cells by a positive selection (CD45 microbeads, Miltenyi Biotec, 130-052-301), incubated with blocking solution (5% normal mouse serum, 5% normal rat serum, and 1% anti-Mouse CD16/CD32) for 30 min on ice, and stained with fluorochrome-conjugated antibody for CD11b (M1/70), CD64 (X54-5/7.1), I-A/I-E (M5/114.15.2), Ly6G (1A8), Ly6C (HK1.4), and CD45 (30-F11, Miltenyi Biotec). All antibodies were from Biolegend, unless otherwise mentioned (clone number in parentheses). Data was acquired with a SH800 flow cytometer (Sony Biotechnology) and analyzed with FlowJo v.10 Software. Mononulear phagocyte cells were gated as CD11b+CD45+Ly6G-I-A/I-E+, as previously described22 (link), and the expression levels of Ly6C and CD64 were analyzed.
Frishberg A., Peshes-Yaloz N., Cohn O., Rosentul D., Steuerman Y., Valadarsky L., Yankovitz G., Mandelboim M., Iraqi F.A., Amit I., Mayo L., Bacharach E, & Gat-Viks I. (2019). Cell composition analysis of bulk genomics using single cell data. Nature methods, 16(4), 327-332.
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2

Isolation and Characterization of Astrocytes

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Mononuclear cells were isolated from the CNS as previously described11 . Astrocytes, monocytes, and microglia were sorted as described before11 and outlined in Supplementary Fig. 1. Isolated CNS cells were stained with fluorochrome-conjugated antibody to CD11b (M1/70, 1:50), CD45 (90, 1:50), Ly6C (HK1.4. 1:100), CD105 (N418, 1:100), CD140a (APA5, 1:100), CD11c (N418, 1:100), F4/80 (BM8, 1:50), O4 (O4, Miltenyi Biotec, 1:10), and CD19 (eBio1D3, 1:100). All antibodies were from eBioscience or BD Pharmingen unless otherwise mentioned. Microglia were sorted as CD11b+ cells with low CD45 expression and low Ly6C (CD11b+CD45lowLy6Clow), inflammatory monocytes were considered as CD45hiCD11b+Ly6Chi61. Astrocytes were sorted as CD11blowCD45low Ly6ClowCD105lowCD140alowCD11blowF4/80lowO4lowCD19low after the exclusion of lymphocytes, microglia, oligodendrocytes, monocytes (Supplementary Fig. 1). Sorted cells were found to be >85% GFAP+ by FACS analysis (Supplementary Fig. 1). We confirmed that we had isolated a relatively pure population of astrocytes by qPCR analysis of the expression of the astrocyte markers gfap, aldh1l1 and aqp4.
Rothhammer V., Mascanfroni I.D., Bunse L., Takenaka M.C., Kenison J.E., Mayo L., Chao C.C., Patel B., Yan R., Blain M., Alvarez J.I., Kébir H., Anandasabapathy N., Izquierdo G., Jung S., Obholzer N., Pochet N., Clish C.B., Prinz M., Prat A., Antel J, & Quintana F.J. (2016). Type I interferons and microbial metabolites of tryptophan modulate astrocyte activity and CNS inflammation via the aryl hydrocarbon receptor. Nature medicine, 22(6), 586-597.
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3

Isolation and Sorting of CNS Immune Cells

Cited 2 times
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Mononuclear cells were isolated from the CNS as previously described, and astrocytes, monocytes, and microglia were sorted as described before.10 (link),20 (link) Isolated CNS cells were stained with fluorochrome-conjugated antibody to CD11b (M1/70, 1:100), CD45 (90, 1:100), Ly6C1 (HK1.4, 1:100), CD105 (N418, 1:100), CD140a (APA5, 1:100), CD11c (N418, 1:100), F4/80 (BM8, 1:50), O4 (O4, Miltenyi Biotec, 1:10), and CD19 (eBio1D3, 1:100). All antibodies were from eBioscience or BD Pharmingen, unless otherwise mentioned. Microglia were sorted as CD11b+ cells with low CD45 expression and low LY6C1 (CD11b+CD45lowLy6C1low), inflammatory monocytes were considered as CD45hiCD11b+Ly6C1hi. Astrocytes were sorted as CD11blowCD45low Ly6C1lowCD105lowCD140alowCD11blowF4/80lowO4lowCD19low after the exclusion of lymphocytes, microglia, oligodendrocytes, and monocytes. Sorted astrocytes were >85% GFAP+ as determined by fluorescence-activated cell sorting analysis and by quantitative PCR (qPCR) analysis of the expression of the astrocyte markers Gfap, Aldh1l1, and Aqp4.
Rothhammer V., Kenison J.E., Li Z., Tjon E., Takenaka M.C., Chao C.C., Alves de Lima K., Borucki D.M., Kaye J, & Quintana F.J. (2021). Aryl Hydrocarbon Receptor Activation in Astrocytes by Laquinimod Ameliorates Autoimmune Inflammation in the CNS. Neurology® Neuroimmunology & Neuroinflammation, 8(2), e946.
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4

Isolation and Sorting of CNS Immune Cells

Cited 2 times
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Mononuclear cells were isolated from the CNS as previously described and astrocytes, monocytes, and microglia were sorted as described before 10 (link),16 (link),17 (link). Isolated CNS cells were stained with fluorochrome-conjugated antibody to CD11b (M1/70, 1:100), CD45 (90, 1:100), Ly6C1 (HK1.4, 1:100), CD105 (N418, 1:100), CD140a (APA5, 1:100), CD11c (N418, 1:100), F4/80 (BM8, 1:50), O4 (O4, Miltenyi Biotec, 1:10), and CD19 (eBio1D3, 1:100). All antibodies were from eBioscience or BD Pharmingen, unless otherwise mentioned. Microglia were sorted as CD11b+ cells with low CD45 expression and low LY6C1 (CD11b+CD45lowLy6C1low), inflammatory monocytes were considered as CD45hiCD11b+Ly6C1hi. Astrocytes were sorted as CD11blowCD45low Ly6C1lowCD105lowCD140alowCD11blowF4/80lowO4lowCD19low after the exclusion of lymphocytes, microglia, oligodendrocytes, and monocytes.
Rothhammer V., Borucki D.M., Tjon E.C., Takenaka M.C., Chao C.C., Fabregat A.A., de Lima K.A., Vazquez C.G., Hewson P., Staszewski O., Blain M., Healy L., Neziraj T., Borio M., Wheeler M., Dragin L.L., Laplaud D.A., Antel J., Alvarez J.I., Prinz M, & Quintana F.J. (2018). Microglial control of astrocytes in response to microbial metabolites. Nature, 557(7707), 724-728.
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5

Isolation and Characterization of Astrocytes

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Mononuclear cells were isolated from the CNS as previously described11 . Astrocytes, monocytes, and microglia were sorted as described before11 and outlined in Supplementary Fig. 1. Isolated CNS cells were stained with fluorochrome-conjugated antibody to CD11b (M1/70, 1:50), CD45 (90, 1:50), Ly6C (HK1.4. 1:100), CD105 (N418, 1:100), CD140a (APA5, 1:100), CD11c (N418, 1:100), F4/80 (BM8, 1:50), O4 (O4, Miltenyi Biotec, 1:10), and CD19 (eBio1D3, 1:100). All antibodies were from eBioscience or BD Pharmingen unless otherwise mentioned. Microglia were sorted as CD11b+ cells with low CD45 expression and low Ly6C (CD11b+CD45lowLy6Clow), inflammatory monocytes were considered as CD45hiCD11b+Ly6Chi61. Astrocytes were sorted as CD11blowCD45low Ly6ClowCD105lowCD140alowCD11blowF4/80lowO4lowCD19low after the exclusion of lymphocytes, microglia, oligodendrocytes, monocytes (Supplementary Fig. 1). Sorted cells were found to be >85% GFAP+ by FACS analysis (Supplementary Fig. 1). We confirmed that we had isolated a relatively pure population of astrocytes by qPCR analysis of the expression of the astrocyte markers gfap, aldh1l1 and aqp4.
Rothhammer V., Mascanfroni I.D., Bunse L., Takenaka M.C., Kenison J.E., Mayo L., Chao C.C., Patel B., Yan R., Blain M., Alvarez J.I., Kébir H., Anandasabapathy N., Izquierdo G., Jung S., Obholzer N., Pochet N., Clish C.B., Prinz M., Prat A., Antel J, & Quintana F.J. (2016). Type I interferons and microbial metabolites of tryptophan modulate astrocyte activity and CNS inflammation via the aryl hydrocarbon receptor. Nature medicine, 22(6), 586-597.
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6

Isolation and Characterization of Lung Macrophages

Cited 1 time
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To validate the performance of CPM we sorted the population of macrophages from the lungs of various CC mice (Supplementary Table 1). To address this, the lungs were dissociated into single cell suspensions using Miltenyi Biotec lung dissociation kit (130-095-927), according to manufacturer’s instructions. Isolated lung cells were then enriched for CD45+ cells by a positive selection (CD45 microbeads, Miltenyi Biotec, 130-052-301), incubated with blocking solution (5% normal mouse serum, 5% normal rat serum, and 1% anti-Mouse CD16/CD32) for 30 min on ice, and stained with fluorochrome-conjugated antibody for CD11b (M1/70), CD64 (X54-5/7.1), I-A/I-E (M5/114.15.2), Ly6G (1A8), Ly6C (HK1.4), and CD45 (30-F11, Miltenyi Biotec). All antibodies were from Biolegend, unless otherwise mentioned (clone number in parentheses). Data was acquired with a SH800 flow cytometer (Sony Biotechnology) and analyzed with FlowJo v.10 Software. Mononulear phagocyte cells were gated as CD11b+CD45+Ly6G-I-A/I-E+, as previously described22 (link), and the expression levels of Ly6C and CD64 were analyzed.
Frishberg A., Peshes-Yaloz N., Cohn O., Rosentul D., Steuerman Y., Valadarsky L., Yankovitz G., Mandelboim M., Iraqi F.A., Amit I., Mayo L., Bacharach E, & Gat-Viks I. (2019). Cell composition analysis of bulk genomics using single cell data. Nature methods, 16(4), 327-332.
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7

Multiparametric Immune Cell Profiling

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Single‐cell suspensions from PBMCs were stained with mAb against human CD14 (HCD14; BioLegend), human CD206 (DCN228; Miltenyi Biotec), human/mouse CD11b (M1/70.15.11.5; Miltenyi Biotec), human CD1a (HI149; Miltenyi Biotec), human CD1a (HI149; Miltenyi Biotec), human CD86 (REA968; Miltenyi Biotec), human CCR7 (REA108; Miltenyi Biotec), human CD45 RA (T6D11; Miltenyi Biotec), CD4 (A161A1; BioLegend), and human CD103 (REA803; Miltenyi Biotec). After 20‐minutes incubation at 4°C in the dark, cells were washed and resuspended in PBS for the FACS analysis. For each test, cells were analyzed using FACSVerse Flow Cytometer and DIVA software (BD Biosciences).
Paparo L., Nocerino R., Ciaglia E., Di Scala C., De Caro C., Russo R., Trinchese G., Aitoro R., Amoroso A., Bruno C., Di Costanzo M., Passariello A., Messina F., Agangi A., Napolitano M., Voto L., Gatta G.D., Pisapia L., Montella F., Mollica M.P., Calignano A., Puca A, & Berni Canani R. (2020). Butyrate as a bioactive human milk protective component against food allergy. Allergy, 76(5), 1398-1415.
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8

Multiparameter Flow Cytometry Analysis

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Single-cell suspensions from murine bone marrow, spleen, PBMCs and RAW264.7 cells were stained with mAb against mouse CD45 (30F11; Miltenyi Biotec; 1:50), mouse CD11b (M1/70.15.11.5; Miltenyi Biotec; 1:50), mouse Ly-6C (REA796; Miltenyi Biotec; 1:50), mouse CD86 (PO3.3; Miltenyi Biotec; 1:10), mouse F4/80 (BM8; Biolegend; 1:10), mouse CD206 (C068C2; Biolegend; 1:50), mouse Qa-2 (695H1-9-9; SONY; 1:50), mouse CD38 (REA616, Miltenyi Biotec; 1:50), mouse SPiDER- β-Gal (SG03-10; Dojindo Molecular Technologies; 1:500), mouse NK1.1 (REA1162, Miltenyi Biotec; 1:50), mouse CD69 (REA937, Miltenyi Biotec; 1:50), mouse CD3 (REA641, Miltenyi Biotec; 1:10). After 20 min incubation at 4 °C in the dark, cells were washed and resuspended in staining buffer for the FACS analysis as previously established [50 (link)]. Additional staining for Saβ-galactosidase was performed after the above-mentioned staining and incubated for 15 min at 37 °C in the dark without washing before FACS analysis. For each test, cells were analyzed using FACS Verse Flow Cytometer (BD Biosciences).
Ciaglia E., Lopardo V., Montella F., Carrizzo A., Di Pietro P., Malavolta M., Giacconi R., Orlando F., Cattaneo M., Madeddu P., Vecchione C, & Puca A.A. (2022). Transfer of the longevity-associated variant of BPIFB4 gene rejuvenates immune system and vasculature by a reduction of CD38+ macrophages and NAD+ decline. Cell Death & Disease, 13(1), 86.
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