Gr1 apc

Manufactured by Thermo Fisher Scientific

Sourced in United States

Gr1-APC is a fluorescent-labeled antibody that binds to the Gr1 antigen, which is expressed on the surface of granulocytes, including neutrophils, eosinophils, and monocytes. It can be used in flow cytometry applications to identify and quantify these cell types in biological samples.

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Lab products found in correlation

Cd105 apc, by BioLegend (3 mentions) F4 80 apc, by Thermo Fisher Scientific (3 mentions) F4 80 fitc, by Thermo Fisher Scientific (3 mentions) Facscalibur, by BD (3 mentions) Caspase 1 p20, by Adipogen (2 mentions) Cd44 pe, by BD (2 mentions) Cd51 pe, by Thermo Fisher Scientific (2 mentions) Gsdmdc1, by Santa Cruz Biotechnology (2 mentions) Cd3 apc, by BioLegend (2 mentions) Cd11c fitc, by BioLegend (2 mentions) Cd90 apc, by BD (2 mentions) Cd14 fitc, by BioLegend (2 mentions) α tubulin, by Proteintech (2 mentions) Il 1β, by Cell Signaling Technology (2 mentions) Cd11b apc, by BioLegend (2 mentions) Nlrp3, by Adipogen (2 mentions) Fitc sca1, by BioLegend (2 mentions) Cd11b fitc, by Thermo Fisher Scientific (2 mentions) Dextran sulfate sodium (dss), by Merck Group (2 mentions) Cd4 fitc, by Thermo Fisher Scientific (2 mentions)

15 protocols using gr1 apc

Induction of Pyroptosis and IBD in Mice

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Lipopolysaccharides (LPS) (E. coli O111:B4) and nigericin (14K05-MM) used to induce pyroptosis were purchased from Invitrogen. Dextran sodium sulfate (DSS) (#118K7374V) used to induce inflammatory bowel disease (IBD) mouse model were obtained from Sigma-aldrich. Anti-mouse antibodies like NLRP3 (#AG-20B-0014, Adipogen), Caspase1-p20 (#AG-20B-0042, Adipogen), IL-1β (#12426, Cell Signaling Technology), GSDMDC1 (#sc-393656, Santa Cruz Biotechnology), alpha Tubulin (#66031-1-1 g, Proteintech), HRP goat anti-mouse IgG antibody (#EK010, Zhuangzhibio), and HRP goat anti-rabbit IgG antibody (#EK020, Zhuangzhibio) were used for immunoblot analysis. The anti-mouse antibodies used for flow cytometry analysis were as follows: APC-CD105 (#MJ7/18, Biolegend), APC-CD90 (#OX-7, BD), PE-CD44 (#IM7, BD, USA), PE-CD51 (#RMV-7, eBioscience), APC-CD3 (#17A2, Biolegend), APC-CD45R/B220 (#RA3-6B2, Biolegend), FITC-CD14 (#Sa14-2, Biolegend), FITC-CD11c (#117306, Biolegend), APC-CD11b (#101212, Biolegend) and APC-Gr1 (#17–5931, eBioscience). The IL-1β Enzyme-Linked Immunosorbent Assay (ELISA) kit was purchased from R＆D Systems (#P16807).

Chen Y., Meng W., Ren G., An N., Zhang J., Liu Z., Wu X., Yin W., Hu X., Liu Z., Feng F, & Chen Y. (2023). NLRP3 inflammasome inhibition of OP9 cells enhance therapy for inflammatory bowel disease. Heliyon, 9(7), e18038.

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Characterizing Mouse Cell Surface Markers

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Antibodies used to characterize mouse cell surface makers by flow cytometry include: mouse PE/Cy7-CD41 (Cat# 25–0411-80) and APC-Gr1 (Cat# 17–5931-82) were purchased from eBioscience. Mouse PE-CD42b antibody (Cat# M040–3) was purchased from Emfret Analytics (Wurzburg, Germany). Antibodies for mouse APC-Ter119 (Cat# 116211), PE/Cy7-CD71 (Cat# 113811), FITC-Mac1 (Cat# 101205), APC/Cy7-cKit (Cat# 105825), FITC-Sca1 (Cat# 108105), PerCP/Cy5.5-FcγR (Cat# 101323), PE-CD150 (Cat# 115903), APC-CD105 (Cat# 120413) and PE-CD34 (Cat# 128609) were purchased from BioLegend (San Diego, CA). PE-Phospho-Stat5 (Cat# 5387) antibody was purchased from Cell Signaling (Danvers, MA). Rabbit polyclonal Von Willebrand Factor (VWF) antibody (Cat# A008229–5) used for immunohistochemistry was purchased from Agilent Technologies (Santa Clara, CA). Recombinant mouse stem cell factor (mSCF, Cat# 250–03), recombinant human EPO (hEPO, Cat# 100–64), recombinant mouse IL-3 (mIL-3, Cat# 213–13), recombinant human TPO (hTPO, Cat# 300–18), recombinant mouse Cxcl1 (mCxcl1, Cat# 250–01), and recombinant mouse Cxcl2 (mCxcl2, Cat# 250–15) were purchased from Peprotech (Rocky Hill, NJ).

Woods B., Chen W., Chiu S., Marinaccio C., Fu C., Gu L., Bulic M., Yang Q., Zouak A., Jia S., Suraneni P.K., Xu K., Levine R.L., Crispino J.D, & Wen Q.J. (2019). Activation of JAK/STAT signaling in megakaryocytes sustains myeloproliferation in vivo. Clinical cancer research : an official journal of the American Association for Cancer Research, 25(19), 5901-5912.

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Characterizing Immune Cell Populations in Mice

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Different cells types in blood, bone marrow and BALF from KO and WT mice were analyzed by flow cytometry. Cells were incubated with FITC-CD11b (eBioscience, San Diego, CA, USA), PC5.5-B220 (BD PharMingen, San Diego, CA, USA) and 780-CD3 (eBioscience) to detect monocytes; with FITC-CD11b and APC-F4/80 (eBioscience) to detect macrophages; with FITC-CD11b and PE-ly6G (eBioscience) to detect neutrophils; with FITC-CD11b, APC-CD11c (eBioscience) and PC5.5-B220 to detect dendritic cells and with FITC-CD11b and APC-Gr-1 (eBioscience) to detect MDSCs. BMMs treated with IL-4 (20 ng/mL) for 48 h were collected, and then incubated with APC-F4/80 (eBioscience) and FITC-CD206 (BioLegend) antibodies at 4 °C for 30 minutes. Cells were analyzed on a flow cytometer (Beckman Coulter, Brea, CA, USA) with Cytexpert software. The experiment was repeated three times.

Li K., Zhang Y., Liang K.Y., Xu S., Zhou X.J., Tan K., Lin J., Bai X.C, & Yang C.L. (2017). Rheb1 deletion in myeloid cells aggravates OVA-induced allergic inflammation in mice. Scientific Reports, 7, 42655.

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Immunoblot and Immunofluorescence Analysis of NLRP3 Inflammasome

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LPS (E. coli O111:B4) and nigericin (14K05-MM) were from Invitrogen. DSS (#118K7374V) were from Sigma-aldrich. Anti-mouse antibodies used for immunoblot analysis were: IL-1β (#12426, Cell Signaling Technologies), NLRP3 (#AG-20B-0014, Adipogen), Caspase-1 p20 (#AG-20B-0042, Adipogen), alpha Tubulin (#66031-1-1g, Proteintech), Caspase-11 p20 (#AG-20B-0061, Adipogen). Anti-mouse antibodies used for immunofluorescent staining analysis were: GSDMDC1 (#sc-393656, Santa Cruz Biotechnology), FITC Goat Anti-Mouse IgG Antibody (L146A, Gene Copoeia), AF647TM Goat Anti-Mouse IgG Antibody (L125A, Gene Copoeia). Anti-mouse antibodies used for flow analysis were: APC-CD45 (#30-F11, Biolegend), APC-Ter119 (#17-5921, eBioscience), PE-CD44 (#IM7, BD, USA), PE-CD51 (#RMV-7, eBioscience), APC-CD90 (#OX-7, BD), APC-CD105 (#MJ7/18, Biolegend), PE-CD146 (#P1H12, eBioscience), PE-CD166 (#105902, R&D), FITC-Sca-1 (#122505, Biolegend), APC-CD3 (#17A2, Biolegend), FITC-CD11c (#117306, Biolegend), APC-CD11b (#101212, Biolegend), APC-Gr1 (#17-5931, eBioscience), FITC-F4/80 (#123108, Biolegend), APC-CD19 (#17-5921, eBioscience), and FITC-CD14 (#Sa14-2, Biolegend).

Chen Y., Qin X., An Q., Yi J., Feng F., Yin D., An N., Liu Z., Weng L., Chen S., Hu X, & Yin W. (2018). Mesenchymal Stromal Cells Directly Promote Inflammation by Canonical NLRP3 and Non-canonical Caspase-11 Inflammasomes. EBioMedicine, 32, 31-42.

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Myeloid Cell Immunophenotyping by Flow

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Bone marrow cells, following red blood cell lysis, or cell lines were stained with FITC-CD11b or APC-Gr1 (eBioscience, CA, USA). Analysis was performed using a PARTEC CyFlow (Munich, Germany) and data were analyzed using FCS EXPRESS.

Li K., Wang F., Cao W.B., Lv X.X., Hua F., Cui B., Yu J.J., Zhang X.W., Shang S., Liu S.S., Yu J.M., Han M.Z., Huang B., Zhang T.T., Li X., Jiang J.D, & Hu Z.W. (2017). TRIB3 Promotes APL Progression through Stabilization of the Oncoprotein PML-RARα and Inhibition of p53-Mediated Senescence. Cancer cell, 31(5).

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FACS Analysis of Myeloid-Derived Suppressor Cells

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Cells were stained with specific antibodies or control isotypes using conventional protocols. The following anti-mouse antibodies were used for FACS analysis of MDSCs: CD11b/PECy5, Gr1/PE, Gr1/APC, CD62L/FITC, F4/80/FITC, CD11c/PE, and MHC CI OVA peptide/Biotin from eBiosciences (San Diego, CA); as well as CD124/PE, CD40/FITC, CD86/FITC, I^A-I^E/FITC, Ly6C/FITC and Ly6G/PE that were purchased from BD Biosciences (San Jose, CA). Lymphocytes characterization was performed with the next specific antibodies: CD8/PE and CD8/Biotin, CD69/PECy5, CD62L/FITC, CD247/PE, CD4/PE, CD4/PECy5, CD25/PECy5, Foxp3/Biotin, IFN-γ/FITC, IL-4/PE and IL-17/PE, all from eBiosciences. FITC-conjugated Streptavidin was purchased also from eBiosciences. In all cases, cells were previously incubated with anti-mouse FcγR 2.4G2 ascites (HB-197; ATCC) to reduce the non-specific binding. For intracellular staining, cells were fixed and permeabilized, according to the manufacturer’s protocol, with either Foxp3 Staining Buffer Set or IC Fixation and 10X Permeabilization Buffers from eBiosciences. Cells were acquired using both FACScan (BD Biosciences) and Gallios (Beckman Coulter, Miami, FL) flow cytometers and analyzed with Kaluza 1.2 (Beckman Coulter) and FlowJo 5.7.2 (Tree Star Inc., USA) softwares.

Fernández A., Oliver L., Alvarez R., Hernández A., Raymond J., Fernández L.E, & Mesa C. (2014). Very small size proteoliposomes abrogate cross-presentation of tumor antigens by myeloid-derived suppressor cells and induce their differentiation to dendritic cells. Journal for Immunotherapy of Cancer, 2, 5.

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Flow Cytometry Analysis of Immune Cells

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Stained cells were analyzed using a BD LSR II-SORP system (Becton Dickinson) and analyzed with FlowJo software (Tree Star). The following mAbs were used: CD45-PerCPCy5, CD45-APC, CD8-APC, anti-IFNγ-PECy7 and anti-IL-4-FITC (BD Biosciences), Gr1-APC, CD11c-PECy7, CD11b-PB, F4/80-APC (eBiosciences), 1A8-APCCy7, Ly6C-FITC, 1A8-APCCy7, CD4-AF700. DAPI and AnV-FITC (BioLegends). In vivo bioluminescence imaging of MPO activity was quantified through injection of 200mg/kg luminol (Carbosynth) i.p. 10 minutes before luminescence acquisition. Photon emission was acquired for 10 minutes using a Xenogen IVIS Imaging system.

Hurrell B.P., Schuster S., Grün E., Coutaz M., Williams R.A., Held W., Malissen B., Malissen M., Yousefi S., Simon H.U., Müller A.J, & Tacchini-Cottier F. (2015). Rapid Sequestration of Leishmania mexicana by Neutrophils Contributes to the Development of Chronic Lesion. PLoS Pathogens, 11(5), e1004929.

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Lung Inflammatory Cell Profiling by Flow Cytometry

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The composition of lung inflammatory cells was determined by flow cytometry (BD FacsCalibur Milan, Italy) using the following antibodies: CD11c-FITC, CD11c-APC, CD11b-PeCy5.5, Gr1-APC, CD3-PeCy5.5, CD4-FITC, CD25-PE, FoxP3-PeCy5.5, F4/80-PE, CD169-FITC, Arg I-PerCp (eBioscience, CA, USA). BAL-derived macrophages were stained for Tetramethyl rhodamine Esther (TMRE, 5 nM, Life Technologies, Monza, Italy) to measure mitochondrial membrane potential alterations. In another set of experiments BAL-derived macrophages were stained for MitoSOX Mitochondrial Superoxide Indicator as indicated in the manufacturer's guide (Life Technologies, USA).

Terlizzi M., Colarusso C., Popolo A., Pinto A, & Sorrentino R. (2016). IL-1α and IL-1β-producing macrophages populate lung tumor lesions in mice. Oncotarget, 7(36), 58181-58192.

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Multicolor Flow Cytometric Analysis of Immune Cell Subsets

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Three-color immunostaining of cell surface and intracellular markers was performed utilizing the Accuri C6 flow cytometer. Isolated splenocytes were stained for CD11b-PE, Gr1-APC, and F480-FITC to evaluate MDSCs; CD3-PE, CD8-FITC, and Granzyme-B-PECy7 (intracellular) to evaluate CD8 cells; and CD4-APC, CD25-FITC, and FoxP3-PE (intracellular) to evaluate T_reg cells (eBio-science). Before intracellular staining, cells were fixed and permeabilized using the FoxP3 Fix/Perm Buffer Set solution (eBioscience). Tumors from mice were morselized into a single cell suspension to evaluate tumor-infiltrating lymphocytes and probed using APC-conjugated anti-mouse antibody directed against CD4 with isotype control (mouse IgG) and PE-conjugated anti-mouse mAb directed against CD8 with isotype control (mouse IgG1) (eBioscience).

Downs-Canner S., Magge D., Ravindranathan R., O’Malley M.E., Francis L., Liu Z., Guo Z.S., Obermajer N, & Bartlett D.L. (2015). Complement Inhibition: A Novel Form of Immunotherapy for Colon Cancer. Annals of surgical oncology, 23(2), 655-662.

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Immunophenotyping of Hematopoietic Cells

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Hematopoietic cells were collected from the bone marrow and peripheral blood of the normal and diseased mice. Erythrocytes were lysed in NH₄Cl red blood cell lysis buffer (pH 7.4). The cells were washed with PBS and stained with Gr-1-APC for neutrophils, B220-PE for lymphocytes, F480-APC for macrophages and Sca-1/c-Kit/Lin for HSC purchased from Ebioscience Inc or cell signaling. After staining, the cells were washed once with PBS and subjected to FACS analysis. Cells were analyzed with FACS calibur or LSRII (Becton Dickinson). Influx cell sorter (Becton Dickinson) was used for sorting of GFP positive cells from cultured myeloid progenitors.

Mukherjee K., Sha X., Magimaidas A., Maifrede S., Skorski T., Bhatia R., Hoffman B, & Liebermann D.A. (2017). Gadd45a deficiency accelerates BCR-ABL driven chronic myelogenous leukemia. Oncotarget, 8(7), 10809-10821.

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