Anti ly6g

Manufactured by BD

Sourced in United States, United Kingdom

Anti-Ly6G is a laboratory reagent used for the detection and analysis of the Ly6G antigen. Ly6G is a cell surface marker expressed on mature granulocytes, such as neutrophils. Anti-Ly6G can be used in various immunological techniques, such as flow cytometry, to identify and quantify Ly6G-positive cells.

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Lab products found in correlation

Anti cd11b, by BD (25 mentions) Anti cd45, by BD (13 mentions) Anti cd4, by BD (13 mentions) Anti ly6c, by BD (13 mentions) Anti f4 80, by BD (9 mentions) Anti cd3, by BD (9 mentions) Anti cd11c, by BD (8 mentions) Anti cd11b, by BioLegend (8 mentions) Anti cd45, by BioLegend (8 mentions) Anti cd11b, by Thermo Fisher Scientific (7 mentions) Anti cd8, by BD (7 mentions) Anti f4 80, by Thermo Fisher Scientific (6 mentions) Anti siglecf, by BD (4 mentions) Anti gr 1, by BD (4 mentions) Anti f4 80, by Bio-Rad (4 mentions) Anti cd8a, by BD (4 mentions) Anti cd3e, by BD (4 mentions) Anti cd19, by BD (3 mentions) Anti ifn γ, by BD (3 mentions) Anti il 17a, by Thermo Fisher Scientific (3 mentions)

Close spelling variants of this product

Anti-Ly6G-PE (17 citations) Anti-Ly6G (clone 1A8, (17 citations) FITC-conjugated anti-Ly6G (7 citations) Anti-Ly6G-APC (7 citations) APC-conjugated anti-Ly6G (6 citations) Anti-Ly6G PerCP (3 citations) FITC anti-mouse Ly6G (3 citations) Purified Rat anti-mouse Ly6G (clone 1A8) (3 citations) Anti-Ly6g PerCP-Cy5.5 (3 citations) Anti-mouse Ly6G (1A8, (3 citations)

64 protocols using anti ly6g

Purification of Kupffer Cells and Inflammatory Monocytes

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To purify KCs, Ly6C^hi and Ly6C^low IMs, liver NPCs were incubated with normal rat serum (Sigma) and anti-mouse FcγRII/III (Becton Dickinson, Franklin Lakes, NJ, USA) to minimize nonspecific antibody binding. Subsequently, the cells were stained with anti-CD45, anti-Ly6C, anti-Ly6G, anti-CD19, anti-SiglecF (Becton Dickinson) and anti-F4/80, anti-CD11b, anti-NK1.1 and anti-CD3 (eBioscience, San Diego, CA, USA), and sorted using a BD FACSAria II Cell Sorter (BD Bioscience, San Jose, CA, USA).

Marentette J.O., Wang M., Michel C.R., Powell R., Zhang X., Reisdorph N., Fritz K.S, & Ju C. (2019). Multi-omics Analysis of Liver Infiltrating Macrophages Following Ethanol Consumption. Scientific Reports, 9, 7776.

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Immunophenotyping of Spleen and Peritoneal Cells

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After cells counting from the peritoneal cavity and spleen, 10⁶ cells/mL of each were resuspended in immunophenotyping specific buffer and transferred to a 96-well U-bottom plate. Then, the cells were labeled with specific antibodies and incubated at 4 °C for 15 min. In the marking of spleen cells, two panels were used with the following fluorochrome-conjugated monoclonal antibodies: FITC-conjugated anti-CD3, anti-CD28 conjugated to PE, and anti-CD4 and anti-CD8 conjugated to PerCP. For the cells of the peritoneum, the 05 marking panels were assembled as follows: anti-CD3 and anti-CD14 conjugated with FITC; anti-CD80, anti-CD86, and anti-CD19 conjugated to PE; and anti-CD4, anti-CD8, anti-iNOS, and anti-Ly6G conjugated to PerCP (Becton Dickinson Biosciences, San Jose, CA, EUA).

Dutra I.L., Araújo L.G., Assunção R.G., Lima Y.A., Nascimento J.R., Vale A.A., Alves P.C., Trovão L.O., Santos A.C., Silva R.M., Silva L.A., Maciel M.C., de Sousa E.M., Elias W.P., Nascimento F.R, & Abreu A.G. (2020). Pic-Producing Escherichia coli Induces High Production of Proinflammatory Mediators by the Host Leading to Death by Sepsis. International Journal of Molecular Sciences, 21(6), 2068.

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Isolation and Analysis of Lung Immune Cells

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Lungs were perfused with PBS containing 2% fetal calf serum (FCS), excised and finely minced, followed by enzymatic digestion for 30 min at 37°C in PBS containing 150U/ml collagenase type IV and 20 U/ml DNase type I. Lung homogenates were suspended in a 20% Percoll gradient and centrifuged. Pellets were then washed, and red blood cells lysed. Cells were incubated in RPMI 10% FCS containing Golgi Plug/Golgi Stop for 2 hours at 37°C. Cells were then stained for 30 minutes with fluorescence-conjugated antibodies (Biolegend or Becton Dickinson) diluted in PBS 2% FCS: anti-CD45 (AF700-conjugated), anti-Ly6G (FITC-conjugated), anti-CD11b (PerCpCy5.5-conjugated), anti-SiglecF (APCCy7-conjugated), anti-CD11c (FITC-conjugated), anti-MHCII (PerCpCy5.5-conjugated), anti-F4/80 (PeCy7-conjugated), anti-CD4 (FITC-conjugated), anti-CD8 (APCCy7-conjugated), anti-B220 (PerCpCy5.5-conjugated) and anti-PD-L1 (PeCy7-conjugated). Cells were washed and fixed, permeabilized, and stained with PE-conjugated antibodies against pro-IL-1β (Thermofisher) or IL-17A (Biolegend) and analyzed on a BD Aria cell sorter. Flow cytometry analyses were performed using the Diva software.

Hassane M., Rahal Z., Karaoghlanian N., Zhang J., Sinjab A., Wong J.W., Lu W., Scheet P., Lee J.J., Raso M.G., Solis L.M., Fujimoto J., Chami H., Shihadeh A.L, & Kadara H. (2022). Chronic exposure to waterpipe smoke elicits immunomodulatory and carcinogenic effects in the lung. Cancer prevention research (Philadelphia, Pa.), 15(7), 423-434.

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Immune Cell Quantification in Tissues

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Tissues were stained with anti-Ly6G (Becton Dickinson, New Jersey, USA), anti-CD3 (Abcam, Cambridge, United Kingdom) or anti-Reg3β (R&D Systems, Minneapolis, Minnesota, USA) antibodies and mounted with ProLong Gold Antifade with DAPI (Life Technologies, Carlsbad, California, USA). Pictures were then taken for each tissue section and positively stained cells counted from at least 4 tissues section.

Sham H.P., Bazett M., Bosiljcic M., Yang H., Luk B., Law H.T., Morampudi V., Yu H.B., Pankovich J., Sutcliffe S., Bressler B., Marshall J.K., Fedorak R.N., Chen J., Jones M., Gunn H., Kalyan S, & Vallance B.A. (2018). Immune Stimulation Using a Gut Microbe-Based Immunotherapy Reduces Disease Pathology and Improves Barrier Function in Ulcerative Colitis. Frontiers in Immunology, 9, 2211.

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Murine BM Macrophage Polarization

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Murine BM cells were cultured in the presence of M-CSF and different doses of LPS as described above. In some experiments, control or IRF5 siRNA (30 pmol, Life Technologies) was also added to cell cultures. After 5 days, cells were harvested and stained with anti-CD11b, anti-Ly6C, anti-Ly6G, anti-CCR5, anti-TLR4, and anti-CD14 antibodies (BioLegend). Propidium iodide (PI) was also added to determine the cell viability. To detect the production of IL-12, BM cells cultured for 5 days were treated with PMA (20 ng/ml), ionomycin (1 μg/ml), and GolgiStopTM protein transport inhibitor (BD Biosciences) for 4 h, and then stained with anti-Ly6C, anti-Ly6G, and anti-CD11b antibodies. After fixation and permeabilization using Cytofix/CytopermTM kit (BD Biosciences), cells were stained with anti-IL-12 antibody (BD Biosciences). The cell phenotype was then analyzed by flow cytometer. The data were processed by FACSDiva or Flow Jo.

Yuan R., Geng S, & Li L. (2016). Molecular Mechanisms That Underlie the Dynamic Adaptation of Innate Monocyte Memory to Varying Stimulant Strength of TLR Ligands. Frontiers in Immunology, 7, 497.

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Isolation of Interscapular Brown Adipose Tissue Cells

Cited 1 time

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Interscapular BAT was dissected from the mice gavaged with Bacteroides or vehicle for 12 weeks and processed for cell isolation as described previously (Hagberg et al., 2018 (link)). Briefly, 100 mg of BAT was minced carefully using scissors for 3 min. The minced tissue was digested in a 37°C thermal shaker (#0003637; TAITEC, Saitama, Japan) for 30 min with Collagenase Type 1 (#LS004196; Worthington Biochemical Corp., Lakewood, NJ) prepared in 10 mL PBS. The samples were then centrifuged for 2 min at 20 × g, and the pellets (stromal vascular fraction) were washed 2 times with 10 mL PBS containing 2% bovine serum albumin and incubated with an anti-CD16/CD32 antibody (#553142; BD Biosciences) to block Fc receptors. This was followed by staining with the following antibodies: anti-CD45 (#557659; BD Biosciences), anti-F4/80 (#565411; BD Biosciences), anti-CD11b (#563402; BD Biosciences), and anti-Ly6G (#560602; BD Biosciences). Samples incubated with the isotype-matched antibodies were used as controls. Flow cytometric analysis was performed on an Attune acoustic focusing cytometer (Life Technologies, Grand Island, NY) and by using FlowJo software (Tree Star, Inc., Ashland, OR).

Yoshida N., Yamashita T., Osone T., Hosooka T., Shinohara M., Kitahama S., Sasaki K., Sasaki D., Yoneshiro T., Suzuki T., Emoto T., Saito Y., Ozawa G., Hirota Y., Kitaura Y., Shimomura Y., Okamatsu-Ogura Y., Saito M., Kondo A., Kajimura S., Inagaki T., Ogawa W., Yamada T, & Hirata K.I. (2021). Bacteroides spp. promotes branched-chain amino acid catabolism in brown fat and inhibits obesity. iScience, 24(11), 103342.

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Immunohistochemical Analysis of Liver Inflammation

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Liver tissues were fixed in 4% neutral buffered formalin and embedded in paraffin. The paraffin sections were deparaffinized by baking and then dehydrated using xylene and ethanol. After blocking with 1% bovine serum albumin (BSA) for 1 h, the sections were incubated with anti-CD11b (1:100, BD Biosciences, USA), anti-Ly6G (1:100; BD Biosciences), anti-H3cit (1:400; CST, USA), anti-MPO (1:100; Abcam, UK), and anti-N-WASP (1:100; Abcam) antibodies overnight at 4°C. Cells were fixed in 4% paraformaldehyde for 25 min at room temperature and then permeabilized with 0.2% Triton X-100 in PBS for 10 min. The cells were blocked in PBS with 2% BSA for 30 min at room temperature and then incubated with anti-H3Cit (1:400; CST), anti-N-WASP (1:100; Abcam), or anti-MPO (1:100; Abcam) antibodies overnight at 4°C. The sections and cells were then incubated with the indicated Alexa Fluor-conjugated secondary antibodies. DAPI was used to detect nuclei. Between all steps, samples were washed three times in PBS for 5 min each. The sections were visualized using an LSM 800 laser scanning confocal microscope (Zeiss, Germany).

Wang L., Zhu Z., Liao Y., Zhang L., Yu Z., Yang R., Wu J., Wu Z, & Sun X. (2022). Host liver-derived extracellular vesicles deliver miR-142a-3p induces neutrophil extracellular traps via targeting WASL to block the development of Schistosoma japonicum. Molecular Therapy, 30(5), 2092-2107.

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Tumor Infiltrating Lymphocyte Isolation

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Tumors were sufficiently chopped using scalpels and then digested with 0.2 mg/mL collagenase intravenous and 0.1 mg/mL DNase I at 37°C for 1 hour, and then passed through a 70 µm strainer to determine the presence of the infiltrating T cell population. Mouse-specific antibodies including anti-CD3, anti-CD4, anti-CD8, anti-CD11b, and anti-Ly6G antibodies were purchased from BD Pharmingen. Anti-IFN-γ and PD-1 antibodies were purchased from eBioscience. The human-specific antibody against PD-L1 was purchased from BD Pharmingen. Major histocompatibility complex (MHC) Class I (H-2Kb) antibody, MHC Class II (I-A/I-E) antibody, human MHC Class I/ HLA-ABC antibody (W6/32) and human MHC Class II/ HLA-DR antibody (LN3) were purchased from eBioscience. Intracellular staining for IFN-γ was performed after stimulation with PMA at 37℃ for 4–6 hours as described in the protocol of the Fixation and Permeabilization Buffer Kit (BD Bioscience). For detection of intracellular IFN-γ, brefeldin A was used to block secretion of cytokines during the last few hours of the stimulation. Cells were then subjected to flow cytometry.

Que Y., Zhang X.L., Liu Z.X., Zhao J.J., Pan Q.Z., Wen X.Z., Xiao W., Xu B.S., Hong D.C., Guo T.H., Shen L.J., Fan W.J., Chen H.Y., Weng D.S., Xu H.R., Zhou P.H., Zhang Y.Z., Niu X.H, & Zhang X. (2021). Frequent amplification of HDAC genes and efficacy of HDAC inhibitor chidamide and PD-1 blockade combination in soft tissue sarcoma. Journal for Immunotherapy of Cancer, 9(2), e001696.

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Multiparametric Flow Cytometry of Tumor-Infiltrating Immune Cells

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Mouse blood was retroorbitally collected in heparinized capillary tubes followed by RBC lysis (BioLegend). Mouse tumors were collected after sacrifice and dissociated using 30% collagenase digestion (30 mins, 37°C). Single cell suspensions in 5% FBS in PBS by passing through 40 μm cell strainer were assessed for viability with Trypan Blue and manually counted, then incubated with Fc receptor blocking solution followed by fluorophore-conjugated antibodies.
For cell surface staining, fluorescence-labeled mouse antibodies (including anti-CD45, anti-CD3, anti-CD4, anti-CD8, anti-CD11b, anti-Ly6C, anti-Ly6G, anti-F4/80, anti-CD11c, anti-CD80, anti-CD86, anti-MHCII purchased from BD Biosciences, BioLegend or eBioscience) were added to samples for 30 min at 4 °C in the dark. For further intracellular staining, cells were washed with PBS supplemented with 5% FBS, permeabilized with Permeabilization Buffer (BD Bioscience) and stained with fluorescence-labeled mouse antibodies (including anti-IFNγ, anti-GzmB, and anti-TNFa purchased from BD Biosciences or BioLegend) for 30 min at 4 °C in the dark. Cells were washed and resuspended in 5% FBS in PBS. Flow cytometry was performed on an LSRII (BD Biosciences), and data analyzed using FlowJo Version X (Tree Star).

Ye J., Mills B.N., Zhao T., Han B.J., Murphy J.D., Patel A.P., Johnston C.J., Lord E.M., Belt B.A., Linehan D.C, & Gerber S.A. (2019). Assessing the magnitude of immunogenic cell death following chemotherapy and irradiation reveals a new strategy to treat pancreatic cancer. Cancer immunology research, 8(1), 94-107.

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Immune Cell Profiling in EAE Mice

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Spinal cords and spleens were separated from naive mice and PBS-, shNC-MSC-, or shBecn1-MSC-treated EAE mice after euthanasia. Mononuclear cells were isolated from spinal cords with Percoll (GE healthcare 17-0891-09) or from spleens with Ficoll (Lymphoprep^TM, 1114547). Cells were stained with anti-CD4 (eBioscience, 17-0041-83; BD Biosciences, 553729), anti-CD8 (BD Biosciences, 553035), anti-PTPRC (45-0452-82), anti-LY6G (11-5931-82), anti-ITGAM (BD Biosciences, 553312), anti-IL2RA (12-0251-83), anti-CD69 (11-0691-82), anti-CXCR3 (12-1831-80), and anti-CCR6 antibodies (Biolegend, 129814). For Th1 cell, Th17 cell and Treg cell analysis, cells were stained with surface marker, permeabilized with the Intracellular Fixation and Permeabilization Buffer Set (eBioscience, 88-8824-00), and then stained with anti-IFNG (11-7311-82), anti-IL17A (12-7177-81), and anti-FOXP3 (12-5773-80) antibodies. All these antibodies were purchased from eBioscience, unless marked otherwise. The samples were analyzed by flow cytometry (FACSAria II, BD Biosciences, San Jose, CA USA)

Dang S., Xu H., Xu C., Cai W., Li Q., Cheng Y., Jin M., Wang R.X., Peng Y., Zhang Y., Wu C., He X., Wan B, & Zhang Y. (2014). Autophagy regulates the therapeutic potential of mesenchymal stem cells in experimental autoimmune encephalomyelitis. Autophagy, 10(7), 1301-1315.

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