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Anti cd45 pe

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Anti-CD45-PE is a fluorescently labeled antibody that binds to the CD45 antigen, which is expressed on the surface of most hematopoietic cells. It is commonly used in flow cytometry applications for the identification and analysis of various cell types.

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Lab products found in correlation

Anti cd11b fitc, by Thermo Fisher Scientific (4 mentions) Anti cd3 apc, by Thermo Fisher Scientific (3 mentions) Anti cd90 fitc, by Thermo Fisher Scientific (2 mentions) Anti cd11b pe cy7, by Thermo Fisher Scientific (2 mentions) Anti cd90 pe, by Thermo Fisher Scientific (2 mentions) Anti cd11b fitc, by BioLegend (2 mentions) Anti cd105 pe, by Thermo Fisher Scientific (2 mentions) Facscanto 2, by BD (2 mentions) Facscalibur flow cytometer, by BD (2 mentions) Anti cd34 pe, by Thermo Fisher Scientific (2 mentions) Anti cd14 fitc, by Miltenyi Biotec (2 mentions) Anti cd31 fitc, by Thermo Fisher Scientific (2 mentions) Anti 105 apc, by Thermo Fisher Scientific (2 mentions) Anti cd34 fitc, by Miltenyi Biotec (2 mentions) Attune nxt, by Thermo Fisher Scientific (2 mentions) Anti pdgfrα cd140a apc, by BioLegend (1 mentions) Anti cd34 fitc, by Thermo Fisher Scientific (1 mentions) Moflo xdp, by Beckman Coulter (1 mentions) Anti cd117 apc, by Thermo Fisher Scientific (1 mentions) Anti cd29 fitc, by Thermo Fisher Scientific (1 mentions)

24 protocols using anti cd45 pe

1

Single-cell RNA-seq of EAE and naive mice

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Prior to euthanasia, old and young EAE and naive SJL/J mice were injected intravenously with 3 μg of anti-CD45-PE (eBioscience, Thermo Fisher Scientific) to label blood-derived and blood vessel–adjacent immune cells. Leptomeninges and cortices were dissected and single-cell suspensions were prepared as previously described. Cells from each individual mouse were stained using BioLegend TotalSeq B hashtags (Hashtags 1–4, catalog numbers 155831, 155833, 155835, 155837) and an anti-PE oligonucleotide barcode (BioLegend, catalog 408113). Cells from each compartment (i.e., leptomeninges) were mixed in a 1:1 ratio, resuspended to a concentration of 1300 cells/μL, and submitted for sequencing on the 10x Genomics platform using 5′ chemistry at the Princess Margaret Genomics Centre in Toronto, Ontario, Canada. Data are available in the NCBI Gene Expression Omnibus database (accession number: GSE201568).
Zuo M., Fettig N.M., Bernier L.P., Pössnecker E., Spring S., Pu A., Ma X.I., Lee D.S., Ward L.A., Sharma A., Kuhle J., Sled J.G., Pröbstel A.K., MacVicar B.A., Osborne L.C., Gommerman J.L, & Ramaglia V. (2022). Age-dependent gray matter demyelination is associated with leptomeningeal neutrophil accumulation. JCI Insight, 7(12), e158144.
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2

Isolation and Characterization of Adipose Cell Populations

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Primary SVF from EAT, SAT, BAT was resuspended in PBS with 2% FBS, and incubated with FACS antibodies for 30 min on ice. Cells were initially selected by size on the basis of forward scatter (FSC) and side scatter (SSC), following separated on the basis of cell-surface markers using a flow cytometer MoFlo XDP (Beckman Coulter, Brea, CA). To detect beige progenitors in SVF, primary SVF was separated on the basis of cell-surface markers including anti-PDGFRα (CD140a)-APC (1:50, BioLegend, San Diego, CA) and anti-CD34-FITC (1:100, eBioscience, San Diego, CA). To quantify MCs in adipose tissues, to isolate MCs from SVF, and to remove MCs from SVF, we stained and sorted MCs with anti-CD34-FITC (1:100, eBioscience), anti-CD45-PE (1:500, eBioscience) and MC markers anti-CD117-APC (1:200, eBioscience) and anti-FCεR1-PE-CY7 (1:200, eBioscience). The MC-removed SVF and MCs (1×105) were collected for TPH1 mRNA expressional analysis.
Zhang X., Wang X., Yin H., Zhang L., Feng A., Zhang Q.X., Lin Y., Bao B., Hernandez L.L., Shi G.P, & Liu J. (2019). Functional Inactivation of Mast Cells Enhances Subcutaneous Adipose Tissue Browning in Mice. Cell reports, 28(3), 792-803.e4.
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3

Flow Cytometric Analysis of MSC Markers

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The positive rate for markers of MSCs and IL-37-MSCs was analyzed by using flow cytometric analysis. In brief, MSCs were stained with fluorescent antibodies, including anti-CD29-FITC, anti-CD45-PE, anti-CD79a-PE, and anti-CD90-FITC (eBioscience, San Diego, USA), according to the manufacturer's instruction. The percentages of various markers of MSCs were analyzed using the FlowJo software.
Kong D., Hu Y., Li X., Yu D., Li H., Zhao Y., Qin Y., Jin W., Zhang B., Wang B., Wang H., Li G, & Wang H. (2020). IL-37 Gene Modification Enhances the Protective Effects of Mesenchymal Stromal Cells on Intestinal Ischemia Reperfusion Injury. Stem Cells International, 2020, 8883636.
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4

Isolation of Microglia from Mouse Brain

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Mice were perfused with PBS containing 5 IU/ml heparin. Isolated brains were stored on ice in HBSS with 45% glucose and HEPES and subsequently minced with a scalpel and dissociated in high-glucose DMEM containing collagenase A, FCS, and DNAse I (30 minutes, 37°C). Cells were centrifuged (10 min, 400g, 4˚C) and resuspended in 5 ml of 25% Percoll overlaid with 3 ml of ice-cold PBS. Microglial cells were obtained as a pellet after centrifugation (30 min, 800g, 4°C), and resuspended in FACS buffer (0.5% BSA, 2mM EDTA in PBS). Microglia were stained (30 min, on ice, in the dark) using anti-CD45-PE (1:800; clone 30-F11, eBioscience), anti-CD11b-APC-Cy7 (1:400; clone M1/70, BD Biosciences), and Fc receptor blocking antibody CD16/CD32 (1:400; clone 2.4G2, BD Biosciences). Microglia were sorted as CD45int CD11b+ cells with an Aria II (BD Biosciences) to >90% purity.
Srinivasan S., Kancheva D., De Ren S., Saito T., Jans M., Boone F., Vandendriessche C., Paesmans I., Maurin H., Vandenbroucke R.E., Hoste E., Voet S., Scheyltjens I., Pavie B., Lippens S., Schwabenland M., Prinz M., Saido T., Bottelbergs A., Movahedi K., Lamkanfi M, & van Loo G. (2024). Inflammasome signaling is dispensable for ß-amyloid-induced neuropathology in preclinical models of Alzheimer’s disease. Frontiers in Immunology, 15, 1323409.
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5

Multi-Parametric Flow Cytometry Analysis

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ENU (CAS no. 759–73-9) and EMS (CAS no. 62–50-0), heparin sodium, low melting point agarose, Triton X-100, dimethyl sulfoxide were purchased from Sigma-Aldrich (Shanghai, China). Anti-CD59-APC and anti-CD71-FITC were obtained from BD Biosciences (San Jose, CA, USA). Anti-CD61-PE and anti-CD45-PE were purchased from eBiosciences (San Diego, CA, USA). DRAQ5 was provided by Abcam (Cambridge, UK). SYTO13 and propidium iodide (PI) were supplied by Invitrogen (Thermo Fisher Scientific, Inc., Waltham, MA, USA). Phosphate-buffered saline (PBS) and Hanks balanced salt mixture (HBSS) and fetal bovine serum (FBS) were purchased from Gibco, Thermo Fisher Scientific (Waltham, MA, USA). Normal melting point agarose and ethylenediaminetetraacetic acid (EDTA) and Tris base were purchased from Amresco (Shanghai, China).
Zhu X., Huo J., Zeng Z., Liu Y., Li R., Chen Y., Zhang L, & Chen J. (2022). Determination of potential thresholds for N-ethyl-N-nitrosourea and ethyl methanesulfonate based on a multi-endpoint genotoxicity assessment platform in rats. Environmental Science and Pollution Research International, 29(56), 85128-85142.
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6

Mouse Liver Cell Isolation and FACS

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Dissected mouse livers were minced with a scalpel on ice and incubated at 37 ˚C in RPMI-1640 media containing 0.05% collagenase/dispase (11097113001; Roche) and 0.01% trypsin inhibitor (T7659; Sigma). Cell suspension was filtered through a 70 μm cell strainer into PBS and residual red blood cells lysed using red cell lysis buffer (11814389001; Roche). Cell suspensions at a concentration of 1 × 107 cells/ml were the incubated for 45 min at 4 ˚C in the presence of anti-CD11b PE-Cy7 (25-0112-82, eBioscience), anti-Gr1 PE (12-5931-82; eBioscience), anti-CD45 PE (12-0451-82; eBioscience), anti-CXCR2-APC (149305; Biolegend), and Mouse BD Fc Block (553141, BD Bioscience). FACS analysis was performed using a FACSCalibur flow cytometer (BD Biosciences) and analyzed with FlowJo software version 7.2.5 (Tree Star, Ashland, OR).
Yuzhalin A.E., Gordon-Weeks A.N., Tognoli M.L., Jones K., Markelc B., Konietzny R., Fischer R., Muth A., O’Neill E., Thompson P.R., Venables P.J., Kessler B.M., Lim S.Y, & Muschel R.J. (2018). Colorectal cancer liver metastatic growth depends on PAD4-driven citrullination of the extracellular matrix. Nature Communications, 9, 4783.
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7

Quantitative Analysis of Human AAT

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Serum from nongrafted hAAT-treated mice was collected using a designated microvette (Fisher Scientific, Waltham, MA). Circulating hAAT levels were detected using species-specific ELISA for human AAT (Immunologic Consultants Laboratory, Inc.). Membrane-associated hAAT was determined by flow cytometry of thioglycolate-elicited peritoneal cell lavages using anti-hAAT-FITC (Bethyl Laboratories, Inc., Montgomery, TX) and anti-CD45-PE (eBioscience) antibodies. Peritoneal macrophages were pulsed with Glassia for indicated time points and lyzed, and hAAT content was depicted by Western blot analysis using goat anti-human AAT (Bethyl Laboratories, Inc.) and mouse anti-β-actin (MP Biomedicals, Santa Ana, CA) antibodies.
Baranovski B.M., Ozeri E., Shahaf G., Ochayon D.E., Schuster R., Bahar N., Kalay N., Cal P., Mizrahi M.I., Nisim O., Strauss P., Schenker E, & Lewis E.C. (2016). Exploration of α1-Antitrypsin Treatment Protocol for Islet Transplantation: Dosing Plan and Route of Administration. The Journal of Pharmacology and Experimental Therapeutics, 359(3), 482-490.
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8

Characterization of ADSC Phenotype

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The cell surface markers were analyzed with flow cytometry. The primary ADSCs at passage 3 (n = 3), and the ADSC-K4DT (n = 3) and ADSC-K4D (n = 3) cells were analyzed at low (approximately PDL 20), intermediate (approximately PDL 50), and high (approximately PDL 100) PDs. The cells were washed with FACS buffer (PBS containing 2% FBS), and the Fc receptors were blocked with canine Fc receptor binding inhibitor (Thermo Fisher Scientific). Then, the cells were incubated with the following phycoerythrin (PE)-conjugated antibodies: anti-CD29-PE (clone: TS2/16; BioLegend, San Diego, CA, USA), anti-CD34-PE (clone: 1H6; R&D Systems, Minneapolis, MN, USA), anti-CD44-PE (clone: IM7; BioLegend), anti-CD45-PE (clone: YKIX716.13; eBioscience, San Diego, CA, USA), anti-CD90-PE (clone: YKIX337.217: eBioscience), and anti-HLA-DR-PE (clone: G46-6: BD Biosciences, Franklin Lake, NJ, USA). All the cells at each passage were examined in triplicate.
Yasumura Y., Teshima T., Nagashima T., Takano T., Michishita M., Taira Y., Suzuki R, & Matsumoto H. (2023). Immortalized Canine Adipose-Derived Mesenchymal Stem Cells as a Novel Candidate Cell Source for Mesenchymal Stem Cell Therapy. International Journal of Molecular Sciences, 24(3), 2250.
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9

Neutrophil Isolation and Characterization

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Migrated mouse neutrophils were collected from the apical compartment following 2h migration. The cell suspension was blocked with fetal bovine serum and anti-CD16/CD32 (1:200, BD Biosciences; San Jose, CA) for 20 min on ice. Suspension was then stained with anti-CD45-PE (eBioscience), and anti-Ly6G-APC (eBioscience), and washed. Data was collected on a Accuri C6 flow cytometer (BD Biosciences), and analyzed with FlowJo analysis software (FlowJo; Ashland, OR).
Pazos M.A., Pirzai W., Yonker L.M., Morisseau C., Gronert K, & Hurley B.P. (2014). Distinct cellular sources of Hepoxilin A3 and Leukotriene B4 are used to coordinate bacterial-induced neutrophil transepithelial migration. Journal of immunology (Baltimore, Md. : 1950), 194(3), 1304-1315.
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10

Isolation and Characterization of BMMSCs from OVX Mice

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BMMSCs were isolated from the femurs and tibias of sham-operated mice or OVX mice treated with PBS, TDNMCP-1, TDNFasL, or TDNs. Femurs and tibias were dissected out and cleaned of connective tissue. Cells were flushed out from long bones by a syringe with α-MEM supplemented with 20% FBS, 2 mM l-glutamine (Sigma-Aldrich, USA), and 1% penicillin/streptomycin (Invitrogen, USA). Single-cell suspensions were seeded in dishes and initially maintained in an atmosphere of 5% CO2 at 37 °C. The medium was changed every 3 days. After reaching 80% confluence, cells were passaged using 0.25% trypsin.
Cell surface markers of BMMSCs were detected by flow cytometry analysis. Cells were harvested and washed in PBS. Then, the single-cell suspension was incubated with mouse anti-CD105-APC (BioLegend, 120413), anti-CD11b-FITC (BioLegend, 101206), anti-CD45-PE (eBioscience, 12-0451), anti-CD29-APC (eBioscience, 17-0291-82) and anti-CD34-PE (BioLegend, 119307) antibodies. Finally, cells were washed twice in PBS, subjected to flow cytometry analysis with a flow cytometer (Cytomics FC 500; Beckman-Coulter) and analyzed with FlowJo_V10 software.
Yang X., Zhou F., Yuan P., Dou G., Liu X., Liu S., Wang X., Jin R., Dong Y., Zhou J., Lv Y., Deng Z., Liu S., Chen X., Han Y, & Jin Y. (2021). T cell-depleting nanoparticles ameliorate bone loss by reducing activated T cells and regulating the Treg/Th17 balance. Bioactive Materials, 6(10), 3150-3163.
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