The following antibodies were purchased from BioLegend, eBioscience, or BD Biosciences, and used for flow cytometry or enrichment of specific cell types: FITC-Sirpα, PE/Cy7-Sirpα PerCP/Cy5.5-B220, PE/Cy7-CD11c, Brilliant Violet 605-CD11c, Pacific Blue-IA/IE, FITC-YAe, PerCP/Cy5.5-CD45.1, APC-CD45.2, PE/Cy7-CD86, APC-CD86, PE-PD-L1, APC-CD200, PE/Cy7-CD40, PE-Foxp3, Pacific Blue-Foxp3, PerCP/Cy5.5-Thy1.1, APC-TCRβ, FITC-CD8α, Alexa700-CD8α, Alexa700-CD4, Brilliant Violet 605-CD4, Brilliant Violet 605-CD25, APC-CD25, PE/Cy7-CD3, PE/Cy7-CD24, Propidium iodide solution, Fixable Viability Dye eFluor780, Biotin-CD8β, Biotin-CD4, Biotin-CD25, and Biotin-B220. AlexaFluor647-conjugated OVA and crimson bead (0.02 μm) were purchased from ThermoFisher Scientific. Streptavidin-RapidSpheres isolation kit and CD11c-MicroBeads were purchased from STEMCELL technologies and Miltenyi Biotec, respectively.
Cd24 pe cy7
Manufactured by BD
Sourced in United States, Belgium
The CD24-PE-Cy7 is a fluorescently labeled antibody that binds to the CD24 cell surface antigen. It is commonly used in flow cytometry applications to identify and characterize cell populations expressing the CD24 marker.
Automatically generated - may contain errors
Lab products found in correlation
Cd44 fitc, by BD (3 mentions)
B220 pacific blue, by BD (3 mentions)
Cd43 apc, by BD (3 mentions)
Cd25 apc, by BD (2 mentions)
Cd11c pe cy7, by BD (2 mentions)
B220 biotin, by BD (2 mentions)
Foxp3 pe, by BD (2 mentions)
Cd8α fitc, by BD (2 mentions)
Biotin cd25, by BD (2 mentions)
Apc tcr β, by BD (2 mentions)
Fitc sirpα, by BD (2 mentions)
Brilliant violet 605 cd11c, by BD (2 mentions)
Pacific blue ia ie, by BD (2 mentions)
Pacific blue foxp3, by BD (2 mentions)
Percp cy5 5 thy1, by BD (2 mentions)
Alexa700 cd8α, by BD (2 mentions)
Brilliant violet 605 cd4, by BD (2 mentions)
Brilliant violet 605 cd25, by BD (2 mentions)
Biotin cd8β, by BD (2 mentions)
Cd71 pe, by BD (2 mentions)
15 protocols using cd24 pe cy7
1
Multiparameter Immune Cell Analysis
2
Isolation and Characterization of CD44+/CD24- Cells
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A single cell suspension of cells cultured under low attachment conditions on poly-HEMA plates was produced by trituration and/or trypsinization. These cells were incubated at 37°C for 2 h to allow for the recovery of cell surface receptors. Cell surface CD44 and CD24 antigens were stained by incubating with FITC-CD44 (BD Biosciences, Cat.#−555478) PE-Cy7-CD24 (BD Biosciences, Cat.#−561646) antibodies for 1 h on ice. Unstained cells, along with corresponding isotype antibodies (Cat. #−552868 and Cat. #−555742—BD Biosciences) served as the appropriate controls.
3
Immunophenotyping and Protein Analysis
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The following antibodies were purchased from BioLegend, eBioscience, or BD Biosciences and used for flow cytometry or enrichment: FITC-Sirpα, PE/Cy7-Sirpα, PerCP/Cy5.5-B220, APC-EpCAM, PE-CD11c, PE/Cy7-CD11c, Brilliant Violet 605-CD11c, Pacific Blue-IA/IE, FITC-CD8α, Alexa700-CD8α, PE-Vα2, APC-Vα2, APC-Vβ5, PE-Vβ6, PE-Foxp3, Pacific Blue-Foxp3, PerCP/Cy5.5-Thy1.1, APC-TCRβ, PE-CD69, PE/Cy7-CD69, Brilliant Violet 605-CD4, APC-CD25, Brilliant Violet 605-CD25, Brilliant Violet 785-CD25, FITC-CD45RB, APC-ICAM1, PR-CD9, PE-CD81, APC-CD48, PE/Cy7-CD24, APC-CD24, PE-CD71, PE/Cy7-CD45, APC-streptavidin, Biotin-Thy1.2, Biotin-CD8β, Biotin-CD25, and Biotin-B220. Alexa647-CTxB subunit was purchased from Molecular Probes. For Western blotting, the following clones were used: MHCII β chain (Bett; homemade), CD48 (OX-78; Serotec), CD24 (M1/69; BioLegend), actin (A2066; Sigma), CD9 (LMC8; eBioscience), and CD81 (Eat2; BioLegend), and HRP-conjugated secondary antibodies (anti-rat, anti-rabbit, and anti-hamster) were purchased from Jackson ImmunoResearch or BIOSOURCE. Myriocin, MβCD, cholesterol, sphingomyelin, and 4-hydroxy-tamoxifen were purchased from Sigma.
4
Western Blotting and Flow Cytometry Antibodies
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Western blotting antibodies were HMGN1 (Aviva Systems Biology, #ARP38532_P050), HMGN1 (Abcam, #ab5212), mouse HMGN1 (affinity purified rabbit polyclonal)34 (link),35 (link), H3K27me3 (Cell Signaling Technologies, #9733, rabbit polyclonal), total H3 (Cell Signaling Technologies, #9715, rabbit polyclonal), and α-tubulin (Sigma, #T9026, mouse monoclonal). Flow cytometry antibodies were B220-Pacific Blue (BD Pharmingen, #558108, clone RA3-6B2), CD43-APC (BD, #560663, clone S7) or CD43-FITC (BD, #561856, clone S7), CD24-PE-Cy7 (BD, #560536, clone M1/69), BP1-PE (eBiosciences, 12-5891, clone 6C3) or BP1-FITC (eBiosciences, 11-5891, clone 6C3), CD45.1-PE-Cy7 (eBiosciences, 25-0453, clone A20), and CD45.2-APC (eBiosciences, 17-0454, clone 104). ChIP-seq antibodies were H3K27me3 (Cell Signaling Technologies, #9733), H3K4me3 (Abcam, #ab8580), and H3K27ac (Abcam, #ab4729).
5
Western Blotting and Flow Cytometry Antibodies
6
Characterization of hESC-Derived Neural Cells
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hESC-derived neural cultures were washed with PBS, trypsinized with 0.05% trypsin for 1–2 min at room temperature and resuspended into single cells using DMEM/F12 with Glutamax containing 10% FBS. For cell surface staining, cells were incubated with CD184-APC (560936; BD Pharmingen), CD24-PE-Cy7 (561646; BD Pharmingen), CD44-PE (51-9007231; BD Pharmingen) and/or CD271-Alexa fluor 647 or BV421 (560877 or 562562; BD Pharmingen) antibodies in PBS containing 1% bovine serum albumen (BSA) according to manufacturer’s protocol. Stained and unstained cells were analyzed using either a BD LSR-II or LSRFortessa flow cytometry machine (BD Biosciences). Control hES9 hESCs or hESC-derived NPCs were used for mCherry control and IgG-PE-Cy7, IgM-FITC, IgG-BV421 and IgG-APC (BD Pharmingen) were used for isotype controls. Cell sorting was performed using either BD FACSAria II or BD Influx cell sorter. Gating for all sorts was defined by isotype control staining. Flow cytometry data analysis was performed using FlowJo software (Tree Star, Inc.).
7
Immunophenotyping of Cell Surface Markers
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Cells (3×105) were blocked with PBS 1× supplemented with 50% FBS and incubated for 30 min with mouse anti human CD44-Phycoerythrin (555479) and CD24-PE-Cy7 (561646) both from BD Biosciences. Cells were then incubated with 7AAD (559925, BD Biosciences). All acquisitions were performed in a FACS Canto II flow cytometer (Becton Dickinson, Frankin Lakes, NJ, USA). Finally, the analysis of flow cytometry data was performed on 7-AAD negative cells using FlowJo V10 software (Tree Star Inc., Ashland, OR, USA).
8
Flow Cytometry Isolation of Mammary Cancer Cells
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MDA-MB-231, MCF7, and SKBR3 human cancer cell lines were trypsinized to single cells, and MMTV-NDL murine mammary tumors (1.0–1.5 cm in diameter) were harvested and dissociated to single cells as previously described [29 (link),30 (link)] with minor modifications. Cells were suspended at 1 × 107 per mL in staining buffer (PBS with 2% FBS) and incubated for 30 min on ice with antibodies. Cells were washed three times, re-suspended in staining buffer with 1 μg/mL Propidium Iodide (PI), analyzed, and sorted with a FACS Aria II cell sorter (Becton Dickinson, Franklin Lakes, NJ, USA). Sorting schemes were based on previously published studies for human cell lines [1 (link),8 (link)] and primary mouse tissues [33 (link),34 (link)]. Results were analyzed using FlowJo software. Antibodies used were CD24-PE-Cy7 (1:100 dilution, #561646) and CD44-APC (1:100 dilution, #559942; BD Pharmingen, San Diego, CA, USA) for human cells, and CD24-PE (1:200 dilution, #553262; BD Pharmingen), CD49f-APC (1:100 dilution, #313615; Biolegend, San Diego, CA, USA), CD31-PE-Cy7 (1:100, #102417; Biolegend), and CD45-PE-Cy7 (1:100, #103113; Biolegend) for mouse cells.
9
Multiparametric B and T Cell Analysis
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The following antibodies comprising the B cell antibody panel were used: B220-V500, CD19-PerCP Cy5.5, CD23-PE, CD21-APC, CD24-PECy7, CD86-V450, MHCII-FITC, and IgM-Biotin (BD Bioscience, Erembodegem, Belgium). T cells panel consisted of the following antibodies: CD4-PerCP, CD3-AlexaFluor 700, CD62L-V500, CD44-FITC, CD28-PE, and CXCR5-V450 (BD Bioscience, Erembodegem, Belgium). Cells were acquired on a FACS Fortessa machine (BD Immunocytometry system, San Jose, CA, USA) and data was analyzed using Flowjo software (Treestar, Ashland, OR, USA).
10
Phenotypic Analysis of Single Cells
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Single cells were labeled with antibodies for CD24-PeCy7 and CD29-PacBlue, or Annexin V-PacBlue (BD Pharmingen), based on the manufacturer’s data sheet. Lineage cells were excluded using the BD Pharmingen lineage cocktail. Dead cells were excluded by staining with Sytox Red (1:1000).
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