Cd24 pe cf594

Manufactured by BD

CD24 PE-CF594 is a fluorescently-labeled antibody that binds to the CD24 antigen. It is a useful tool for the identification and analysis of CD24-expressing cells in flow cytometry applications.

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CD24-PE (78 citations)

3 protocols using cd24 pe cf594

Comprehensive Immune Cell Profiling

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The following monoclonal antibodies were purchased from eBioscience: CD278 (ICOS)-biotin, CD27-PeCy7, Foxp3-FITC, RORγt-PE, T-bet-PE; or from BD Biosciences: PD1-PECF594, CXCR3-APC, CD24-PECF594, CD25-BB515, CD44-PECy7, CD4-A700, CD8-A700, CD4-PB, CD62L-A700, GATA3-PE, RORγt-PECF594, STAT1 (pY701)-A488, IFNγ-PE, IL-10-APC, IL-17-PerCP-Cy5.5, streptavidin-PECy7. Live/dead fixable near-IR stain (Thermo Fisher) was used to exclude dead cells. For transcription factor staining, cells were stained for surface markers, followed by fixation and permeabilization before nuclear factor staining according to the manufacturer’s protocol (Foxp3 staining buffer set from eBioscience). For cytokine staining, cells were stimulated in media containing phorbol 12-myristate 13-acetate (50 ng/ml, Sigma-Aldrich), ionomycin (250 ng/ml, Sigma-Aldrich), and brefeldin-A (1/100, eBioscience) for 3 hr. After stimulation, cells were stained for surface markers, followed by fixation and permeabilization before intracellular staining according to the manufacturer’s protocol (cytokine staining buffer set from BD Biosciences). For phosphorylation staining, cells were stimulated with IFN-γ (50 ng/ml, PeproTech) for 30 min, fixed with formaldehyde, and permeabilized with methanol before staining. Flow cytometric analysis was performed on a Canto II (BD Biosciences) and analyzed using FlowJo software (Tree Star).

Ajouaou Y., Azouz A., Taquin A., Denanglaire S., Hussein H., Krayem M., Andris F., Moser M., Goriely S, & Leo O. (2022). The oxygen sensor prolyl hydroxylase domain 2 regulates the in vivo suppressive capacity of regulatory T cells. eLife, 11, e70555.

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Immunophenotyping Immortalized B Cells

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Mock- or MCM-treated immortalized B cells or TAB were stained with the following antibodies or matched isotypes and analyzed on a FACS Aria III (BD): CD19 BV711 (clone SJ25C1, 0.06 µg/100 µl, catalog number 563036), CD20 AF700 (clone 2H7, 0.5 µg/100 µl, 560631), CD24 PE-CF594 (clone ML5, 1 µg/100 µl, 562405), CD27 BV421 (clone M-T271, 0.25 µg/100 µl, 562513), CD38 APC (clone HIT2, 0.125 µg/100 µl, 555462), CD138 PE (clone MI15, 0.125 µg/100 µl, 552026), IgD PE-Cy7 (clone IA6-2, 0.125 µg/100 µl, 561314), IgG FITC (clone G18-145, 0.125 µg/100 µl, 555786), IgM BV605 (clone G20-127, 0.5 µg/100 µl, 562977) (all BD biosciences). Live/dead cell exclusion was performed by addition of 7-AAD (5 µg/ml, Calbiochem) prior to acquisition of the samples. Data were analyzed using FlowJo 10.4.2 (FlowJo LLC). The gating strategy is shown in Supplementary Fig. 10.

Griss J., Bauer W., Wagner C., Simon M., Chen M., Grabmeier-Pfistershammer K., Maurer-Granofszky M., Roka F., Penz T., Bock C., Zhang G., Herlyn M., Glatz K., Läubli H., Mertz K.D., Petzelbauer P., Wiesner T., Hartl M., Pickl W.F., Somasundaram R., Steinberger P, & Wagner S.N. (2019). B cells sustain inflammation and predict response to immune checkpoint blockade in human melanoma. Nature Communications, 10, 4186.

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Multiparameter Flow Cytometry Immunophenotyping

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Single-cell suspensions were incubated with human Fc receptor-binding inhibitor (Fc Block, eBioscience) before surface staining with anti-human monoclonal antibodies (mAbs). Fluorescence minus one (FMO) controls with isotype controls were used to define positive signals for flow cytometry or cell sorting. Dead cells were excluded with the Zombie Violet Fixable Viability Kit (Biolegend).
For labeling, cells were resuspended in PBS, 2% FCS (1 to 5 x10 7 cells/500 ml) and incubated with the following mAbs: CD45 A700 (Biolegend, clone HI30), CD34 PB (Biolegend, clone 581), CD38 BV510 (BD Bioscience, clone HIT2), CD117 BV605 (Biolegend, clone 104D2), CD45RA PE (BD Bioscience, clone HIT100), CD7 FITC (Beckman Coulter, clone 8H8.1), CD2 PerCPCy5.5 (Biolegend, clone RPA2.10), CD115 APC (Biolegend, clone 9-4D2-1E4), CD116 APC-vio770 (Miltenyi, clone REA211), CD123 BV786 (BD Bioscience, clone 7G3), CD127 PC5 (Biolegend, clone A019D5), ITGB7 PC7 (eBioscience, clone FIB504), CD10 BV650 (BD bioscience, clone HI10a), CD19 BV711 (BD bioscience, clone HIB19), CD24 PE-CF594 (BD bioscience, clone ML5). Flow cytometry analyses and cell sorting were performed with a BD Fortessa Analyzer or a BD FACSAria IIII sorter (BD Biosciences; (purity R 95%).

Alhaj Hussen K., Vu Manh T.P., Guimiot F., Nelson E., Chabaane E., Delord M., Barbier M., Berthault C., Dulphy N., Alberdi A.J., Burlen-Defranoux O., Socié G., Bories J.C., Larghero J., Vanneaux V., Verhoeyen E., Wirth T., Dalod M., Gluckman J.C., Cumano A, & Canque B. (2017). Molecular and Functional Characterization of Lymphoid Progenitor Subsets Reveals a Bipartite Architecture of Human Lymphopoiesis. Immunity, 47(4).

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