Cd45 pe cy7

Manufactured by Cytek Biosciences

CD45 (PE-cy7) is a fluorescently-labeled antibody that targets the CD45 antigen, which is expressed on the surface of most hematopoietic cells. This reagent can be used in flow cytometry applications to identify and quantify different cell populations.

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2 protocols using cd45 pe cy7

Quantifying Immune Cell Profiles in Intracranial Melanoma

Cited 3 times

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Flow cytometry was used to analyze immune cell populations (12 (link), 13 (link), 44 (link)) in a minimum of three samples in two independent animal studies. Dissected intracranial melanoma tumors were physically dissociated and filtered through a 70-μm cell filter. Single cells were labeled with primary antibody. Cells without primary antibody labeling were used as unlabeled negative controls; fluorescent beads (UltraComp eBeads^™, Invitrogen^™, Carlsbad, CA) were used as positive/calibration controls and to determine compensation between fluorescent channels. Forward- and side-scatter gating identified single cells and viable cell (Ghost Dye^™ Red 780 Viability Dye, 1:100; Tonbo Biosciences, San Diego, CA) exclusion identified live cells. Fluorescence minus one (FMO) methodology was used to determine gating. Flow cytometry was performed on an Attune NxT Flow Cytometer (Thermo Fisher Scientific^™ Inc., Waltham, MA), and compensation matrix computed and data analyzed using FlowJo version 9 software (Ashland, OR) following published flow cytometry guidelines (45 (link)).
Antibodies used were: CD4 (BV510, 1:400, clone RM4–5), CD8 (PerCP-cy5.5, 1:200, clone 53–6.7), FOXP3 (BV421, 1:100, clone MF-14), all acquired from BioLegend^® Inc. (San Diego, CA); and CD45 (PE-cy7, 1:200, clone 30-F11), CD3 (FITC, 1:200, clone 17A2), both acquired from Tonbo Biosciences.

Clark P.A., Sriramaneni R.N., Bates A.M., Jin W.J., Jagodinsky J.C., Hernandez R., Le T., Jeffery J.J., Marsh I.R., Grudzinski J.J., Aluicio-Sarduy E., Barnhart T.E., Anderson B.R., Chakravarty I., Arthur I.S., Kim K., Engle J.W., Bednarz B.P., Weichert J.P, & Morris Z.S. (2021). Low-Dose Radiation Potentiates the Propagation of Anti-Tumor Immunity against Melanoma Tumor in the Brain after In Situ Vaccination at a Tumor outside the Brain. Radiation research, 195(6), 522-540.

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Immunophenotyping of Cultured Cells

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Digested cells were suspended in HBSS at a density of 1–5 × 10⁷ cells/mL and stained for 30 minutes on ice with THY-1-APC (BD Pharmingen) and LNGFR-PE (Miltenyi Biotec) antibodies for sorting. Cultured cells (at three and five passages) were harvested using cell-dissociation buffer (Gibco). Cells (1.0 × 10⁵) were suspended in ice-cold HBSS and stained for 30 minutes on ice with the monoclonal antibodies CD45-PE-Cy7 (Tonbo Biosciences), CD29-PE, CD31-PE-Cy7, CD44-PE, CD105-PE and CD166-PE (BioLegend) for cell surface analysis. Flow cytometric analysis and cell sorting were performed on a triple-laser Moflo system (Beckman Coulter) and data were analyzed using Flowjo software (Tree Star).

Ogata Y., Mabuchi Y., Yoshida M., Suto E.G., Suzuki N., Muneta T., Sekiya I, & Akazawa C. (2015). Purified Human Synovium Mesenchymal Stem Cells as a Good Resource for Cartilage Regeneration. PLoS ONE, 10(6), e0129096.

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