Mac 1 m1 70

Manufactured by BD

The Mac-1 (M1/70) is a lab equipment product designed for general laboratory use. It serves as a multipurpose tool for various tasks within a research or testing environment. The core function of the Mac-1 (M1/70) is to provide a reliable and versatile platform for laboratory operations.

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Lab products found in correlation

B220 ra3 6b2, by BD (3 mentions) Fibrinogen, by Merck Group (2 mentions) Cy3 conjugated anti rat and anti rabbit, by Jackson ImmunoResearch (2 mentions) Cd3 145 2c11, by BD (2 mentions) Gr 1 rb6 8c5, by BD (2 mentions) Cd8 53 6, by BD (2 mentions) Cd4 gk1, by BD (2 mentions) Secondary biotinylated goat anti rat igg, by Vector Laboratories (2 mentions) Ly6g mab, by BD (2 mentions) Abc kit, by Vector Laboratories (2 mentions) Fibronectin, by Merck Group (1 mentions) Fluoromyelin red, by Thermo Fisher Scientific (1 mentions) C kit 2b8, by BD (1 mentions) Cd44 im7, by BD (1 mentions) Cd25 7d4, by Thermo Fisher Scientific (1 mentions) Cd43 s7, by BD (1 mentions) Ter119 ter119, by BD (1 mentions) Cd4 h129, by BD (1 mentions) Cd45.1 a20, by BD (1 mentions) Cd45.2 104, by BD (1 mentions)

8 protocols using mac 1 m1 70

Immunohistochemistry of Cellular Markers

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Immunohistochemistry was performed on 10 μm frozen sections of cold phosphate buffer saline (PBS) perfused tissues as described previously [26 (link)]. Antibodies reactive for the following antigens were used in this study: rat monoclonals reactive to CD31 (MEC13.3), α5 integrin (5H10–27 (MFR5)), CD45 and Mac-1 (M1/70), all from BD Pharmingen (La Jolla, CA); mouse monoclonal to Ki67 from Vector Labs (Burlingame, CA), and rabbit polyclonals reactive to fibronectin (Sigma) and fibrinogen from Millipore (Temecula, CA). Fluoromyelin-red was obtained from Invitrogen. Secondary antibodies used included Cy3-conjugated anti-rat and anti-rabbit from Jackson Immunoresearch, (West Grove, PA) and Alexa Fluor 488-conjugated anti-rat and anti-mouse from Invitrogen (Carlsbad, CA).

Kant R., Halder S.K., Bix G.J, & Milner R. (2019). Absence of endothelial α5β1 integrin triggers early onset of experimental autoimmune encephalomyelitis due to reduced vascular remodeling and compromised vascular integrity. Acta Neuropathologica Communications, 7, 11.

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Hematopoietic Cell Profiling in Mouse Tissues

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Hematopoietic stem and progenitor cell and lineage-positive cell profiling was perform on BM, spleen, and thymus of mice from each genotype. Antibodies (clones) used: CD3 (145-2C11) [BD: #553064], CD4 (H129.19) [BD: #553653], CD8 (53-6.7) [BD: #553033], Gr-1 (RB6-8C5) [BD: #553128], B220 (RA3-6B2) [BD: #553090], Ter119 (Ter119) [BD: #553673], Mac1 (M1/70) [BD: #553311], IL7Rα (A7R34) [eBiosceinces: #12-1271-82], cKit (2B8) [BD: #558163], Sca-1 (E13-161.7) [eBiosceinces: #17-5981-83], CD25 (7D4) [eBiosciences: 13-0252-82], CD44 (IM7) [BD: 559250], CD43 (S7) [BD: 561856], HSA (M1/69) [BD: #553262] and BP-1 (6C3) [BD: #553159], CD4 (GK1.5) [BD: #561830], CD8 (7D4) [eBiosciences: #12-0081-82], B220 (RA3-6B2) [BD: #561880].
Flow cytometric gating strategy is described in Supplementary Fig. 6.

Khattar E., Maung K.Z., Chew C.L., Ghosh A., Mok M.M., Lee P., Zhang J., Chor W.H., Cildir G., Wang C.Q., Mohd-Ismail N.K., Chin D.W., Lee S.C., Yang H., Shin Y.J., Nam D.H., Chen L., Kumar A.P., Deng L.W., Ikawa M., Gunaratne J., Osato M, & Tergaonkar V. (2019). Rap1 regulates hematopoietic stem cell survival and affects oncogenesis and response to chemotherapy. Nature Communications, 10, 5349.

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Bone Marrow and Spleen Cell Isolation

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Bone marrow cells were harvested from femurs and tibiae by crushing the bones with a mortar and pestle. The cell suspension was collected after filtering through 40 μ membrane filter (BD). These cells were used for surface staining after red blood cell (RBC) lysis treatment with Red Blood Cell Lysing Buffer Hybri-Max™ (Sigma). Spleens were crushed between two glass slides and isolated cells were used for surface staining with respective antibodies after RBC lysis. Surface staining was done with B220 (RA3-6132, BD), CD3 (145-2C11, BD), Gr-1 (RB6-8C5, BD), Mac1 (M170, BD), CD45.1 (A20, BD), and CD45.2 (104, BD). After surface staining with respective fluorescent antibodies, polychromatic flow acquisition was performed using a LSR II instrument (BD) at the University of Rochester Flow Cytometry Core. Flow data was analyzed using FlowJo software (version 10.0.7 Treestar, Ashland, OR).

Unnisa Z., Singh K.P., Henry E.C., Donegan C.L., Bennett J.A, & Gasiewicz T.A. (2016). Aryl Hydrocarbon Receptor Deficiency in an Exon 3 Deletion Mouse Model Promotes Hematopoietic Stem Cell Proliferation and Impacts Endosteal Niche Cells. Stem Cells International, 2016, 4536187.

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Isolation of Murine Hematopoietic Stem Cells

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Single-cell suspensions of whole BM cells were labeled by incubation on ice for 15 min using a cocktail of the following antibodies: PE-conjugated anti-mouse CD3ε (145-2C11; BD Biosciences), CD4 (L3T4; BD Biosciences), CD8α (53-6.7; BD Biosciences), B220 (RA3-6B2; BD Biosciences), Mac-1 (M1/70; BD Biosciences), Gr-1 (RB6-8C5; eBioscience), and Ter119 (TER-119, eBioscience); FITC-conjugated anti-mouse Sca-1 (E13-161.7; BD Biosciences); and APC-conjugated anti-mouse c-Kit (2B8; BD Biosciences). After cells were washed with PBS, they were incubated with anti-PE microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany) at 4°C for 15 min. Lin⁺ cells (CD4⁺, CD8α⁺, B220⁺, Mac-1⁺, Gr-1⁺, Ter119⁺ cells) were removed by immunomagnetic selection using a MACS system (Miltenyi Biotec). These negatively selected cells were subjected to FACS using a Moflo XDP cell sorter (Beckman-Coulter) to prepare LSK cells.

Sato F., Miyaoka Y., Miyajima A, & Tanaka M. (2014). Oncostatin M Maintains the Hematopoietic Microenvironment in the Bone Marrow by Modulating Adipogenesis and Osteogenesis. PLoS ONE, 9(12), e116209.

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Quantifying Liver Inflammatory Cells

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Liver sections were stained with hematoxylin and eosin (H&E). The necrotic area was assessed by analyzing at least 10 randomly selected areas per sample, with computer-assisted image analysis with MetaMorph software (Universal Imaging Corporation, Downingtown, PA). Liver macrophages and neutrophils were detected using primary rat anti-mouse F4/80⁺ mAb (Mac-1, M1/70; BD Biosciences, San Jose, CA) and Ly6G⁺ mAb (BD Biosciences, San Diego, CA), respectively. After incubation with secondary biotinylated goat anti-rat IgG (Vector, Burlingame, CA), followed by treatment with immunoperoxidase (ABC Kit, Vector), the average number of positive cells was quantified by analyzing at least 10 random high-power fields (HPF, original magnification × 400) per animal, with Image-Pro Plus software.

Yu M., Zhou M., Li J., Zong R., Yan Y., Kong L., Zhu Q, & Li C. (2022). Notch-activated mesenchymal stromal/stem cells enhance the protective effect against acetaminophen-induced acute liver injury by activating AMPK/SIRT1 pathway. Stem Cell Research & Therapy, 13, 318.

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Isolation and Flow Cytometry Analysis of Immune Cells

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Spleens and thymi were collected in PBS, and cells were obtained by mechanical disruption. Cell suspensions were treated with lysis buffer (0.15 M NH₄Cl, 1 mM NaHCO3, 0.1 mM EDTA) for 5 min at room temperature. Cells were washed and resuspended in FACS buffer for flow cytometry analysis. The following Abs from BD Biosciences were used for flow cytometry: CD4-GK1.5, CD8-53-6.7, CD19-1D3, B220-RA3-6B2, Mac-1–M 1/70, and isotype control. Abs to stain various TCR Vβ regions were also obtained from BD Biosciences. Foxp3⁺ T cells were enumerated using an intracellular staining kit from eBioscience (San Diego, CA). Flow cytometry was performed on a FACScan (BD Biosciences) and analyzed using CellQuest software. Mononuclear cells were gated based on forward and side scatter profiles. Subsequent gating was based on staining with the indicated Abs.

Chowdhary V.R., Dai C., Tilahun A.Y., Hanson J.A., Smart M.K., Grande J.P., Rajagopalan G., Fu S.M, & David C.S. (2015). A Central Role for HLA-DR3 in Anti-Smith Antibody Responses and Glomerulonephritis in a Transgenic Mouse Model of Spontaneous Lupus. Journal of immunology (Baltimore, Md. : 1950), 195(10), 4660-4667.

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Immunohistochemical Characterization of Vascular and Immune Markers

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Immunohistochemistry was performed on 10-μm frozen sections of cold phosphate buffer saline (PBS)-perfused tissues as described previously [33 (link)]. Antibodies reactive for the following antigens were used in this study: rat monoclonals reactive to CD31 (MEC13.3), β4 integrin (346-11A), MHC class II (M5/114.15.2), CD45, Mac-1 (M1/70), all from BD Pharmingen (La Jolla, CA); mouse monoclonal α-SMA-Cy3 conjugate (1A4) from Sigma (St. Louis, MO); rabbit polyclonals reactive to claudin-5 and ZO-1 from Invitrogen (Carlsbad, CA); and fibrinogen from Millipore (Temecula, CA). Secondary antibodies used included Cy3-conjugated anti-rat and anti-rabbit from Jackson Immunoresearch (West Grove, PA) and anti-rat Alexa Fluor 488 and anti-rabbit Alexa Fluor 568 from Invitrogen (Carlsbad, CA).

Welser J.V., Halder S.K., Kant R., Boroujerdi A, & Milner R. (2017). Endothelial α6β4 integrin protects during experimental autoimmune encephalomyelitis-induced neuroinflammation by maintaining vascular integrity and tight junction protein expression. Journal of Neuroinflammation, 14, 217.

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Liver Macrophage and Neutrophil Detection

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Liver macrophages and neutrophils were detected using primary rat anti-mouse CD11b⁺ mAb (Mac-1, M1/70; BD Biosciences, San Jose, CA) or Ly6G⁺ mAb (BD Biosciences, San Diego, CA). After incubation with secondary biotinylated goat anti-rat IgG (Vector, Burlingame, CA), followed by treatment with immunoperoxidase (ABC Kit, Vector), positive cells were counted blindly in ten HPF/section (×400). For immunofluorescence, frozen liver sections were labeled with primary antibodies ATF3 and CD11b (Santa Cruz Biotechnology, CA), and then incubated with secondary Cy3-conjugated AffiniPure donkey anti-goat IgG antibody (Jackson Immunoresearch, PA). The samples were pre-mounted with VECTASHIELD medium with DAPI.

Zhu Q., Wang H., Jiang B., Ni X., Jiang L., Li C., Wang X., Zhang F., Ke B, & Lu L. (2018). Loss of ATF3 exacerbates liver damage through the activation of mTOR/p70S6K/ HIF-1α signaling pathway in liver inflammatory injury. Cell Death & Disease, 9(9), 910.

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