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Apc conjugated cd44

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APC-conjugated CD44 is a fluorescently labeled antibody that binds to the CD44 cell surface receptor. CD44 is a glycoprotein involved in cell-cell interactions, cell adhesion, and migration. The APC (Allophycocyanin) fluorescent dye is conjugated to the CD44 antibody, allowing for detection and analysis of CD44-expressing cells using flow cytometry or other fluorescence-based techniques.

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Lab products found in correlation

Pe conjugated cd133, by Miltenyi Biotec (4 mentions) Facscalibur, by BD (4 mentions) Facsaria, by BD (3 mentions) Cd105, by Miltenyi Biotec (3 mentions) Pe conjugated cd34, by BD (1 mentions) Fitc conjugated cd45, by BD (1 mentions) Pe conjugated cd73, by BD (1 mentions) Af 700 conjugated cd3, by BD (1 mentions) Apc conjugated cd105, by BioLegend (1 mentions) Facsdiva software, by BD (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Pe conjugated cd14, by BD (1 mentions) Abcg2, by R&D Systems (1 mentions) Mal 2 lectin, by Vector Laboratories (1 mentions) Fitc conjugated cd24, by BD (1 mentions) Biotinylated sna lectin, by Vector Laboratories (1 mentions) Facsaria 2 cell sorter, by BD (1 mentions) Rnaprotect cell reagent, by Qiagen (1 mentions) Propidium iodide, by Thermo Fisher Scientific (1 mentions) Annexin 5, by BD (1 mentions)

9 protocols using apc conjugated cd44

1

Multiparameter Profiling of Cancer Cell Subpopulations

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1 × 106 knock-down cells were incubated with BV421-conjugated CD326 (563180, BD, USA), PE-conjugated CD133 (130-080-801, Miltenyi Biotec, Germany), APC-conjugated CD44 (559942, BD) for 30 min at room temperature in the dark. Isotypic IgG and unstained cells were used as negative controls. We set gates for EpCAM+CD133+CD44+ (E/133+44+), EpCAM+CD133−CD44+ (E/44+), EpCAM+CD133+CD44−(E/133+) and EpCAM+CD133−CD44− (E+), and then calculated the proportions of each subpopulation.
Zhang X., Yang L., Lei W., Hou Q., Huang M., Zhou R., Enver T, & Wu S. (2022). Single-cell sequencing reveals CD133+CD44−-originating evolution and novel stemness related variants in human colorectal cancer. eBioMedicine, 82, 104125.
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2

Multiparametric Flow Cytometry Analysis

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FACS analysis and cell sorting were performed using FACS Calibur and FACS Aria machines (Becton Dickinson, Palo Alto, CA), respectively. FACS data were analyzed using FlowJo software (Tree Star, Ashland, OR). Antibodies to the following proteins were used: APC-conjugated CD44 (BD Bioscience, Cat. 559942, dilution 1/40), PE-conjugated CD133 (MACS; Miltenyi Biotech, Sunnyvale, CA, 130-080-081, dilution 1/40), CD34 (MACS; Miltenyi Biotech, Sunnyvale, CA, 30-081-002), CD44 (MACS; Miltenyi Biotech, Sunnyvale, CA, 130-095-180), CD45 (MACS; Miltenyi Biotech, Sunnyvale, CA, 130-080-201), CD73 (MACS; Miltenyi Biotech, Sunnyvale, CA, 130-095-182) and CD105 (MACS; Miltenyi Biotech, Sunnyvale, CA, 130-094-941) The FACS gates were established by staining with an isotype antibody or secondary antibody.
Park S.R., Cho A., Park S.T., Park C.H., Lim S., Jin M., Lee H.Y, & Hong I.S. (2018). Double-edged sword of gonadotropin-releasing hormone (GnRH): A novel role of GnRH in the multiple beneficial functions of endometrial stem cells. Cell Death & Disease, 9(8), 828.
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3

Multiparameter Flow Cytometry Profiling

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Cultured cells (passage 1) were trypsinized and stained with a panel of antibodies for fluorescence-activated cell sorting (FACS) analysis. Approximately, 1 × 105 cells were re-suspended in phosphate buffered saline (PBS) and incubated with IgG block for 5 minutes to block non-specific binding. The following antibodies were used: AF-700 conjugated CD3 (BD BioSciences, USA), PE conjugated CD14 (BD, Immunocytometry, USA), APC conjugated CD19 (BD BioSciences, USA), PE conjugated CD34 (BD, BioSciences, USA), APC conjugated CD44 (BD, Pharmingen, USA), FITC conjugated CD45 (BD Pharmingen, USA), PE conjugated CD73 (BD Pharmingen, USA), AF-700 conjugated CD90 (Biolegend, USA) and APC conjugated CD105 (Biolegend, USA). Cells were stained for 30 minutes at 4°C with the antibodies. After washing, samples were analyzed on a LSR II flow cytometer (BD, USA) and at least 10,000 events were acquired for each population. Data acquisition and analysis were performed using FACS DIVA software (BD Biosciences, USA). Unstained cells were used to establish flow cytometer settings. Debris and cells/particles with auto-fluorescence were removed by using a threshold on the forward scatter.
Choudhery M.S., Badowski M., Muise A., Pierce J, & Harris D.T. (2014). Donor age negatively impacts adipose tissue-derived mesenchymal stem cell expansion and differentiation. Journal of Translational Medicine, 12, 8.
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4

Pancreatic Cancer Cell Immunophenotyping

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Human pancreatic cancer cells were dissociated with trypsin, washed twice with HBSS containing 2% fetal bovine serum and 10 mM HEPES (staining buffer). Cells were stained and incubated on ice for 30 minutes with the following primary antibodies: ABCG2 (R&D Systems Inc, 1:50), FITC-conjugated CD24 (BD Biosciences, 1:100), APC-conjugated CD44 (BD Biosciences, 1:100), biotinylated SNA lectin (Vector Laboratories, 1:100), or MALII lectin (Vector Laboratories, 1:100). After stained with appropriated secondary antibody, the stained cells were analyzed by FACSCalibur (BD Biosciences).
Hsieh C.C., Shyr Y.M., Liao W.Y., Chen T.H., Wang S.E., Lu P.C., Lin P.Y., Chen Y.B., Mao W.Y., Han H.Y., Hsiao M., Yang W.B., Li W.S., Sher Y.P, & Shen C.N. (2016). Elevation of β-galactoside α2,6-sialyltransferase 1 in a fructose-responsive manner promotes pancreatic cancer metastasis. Oncotarget, 8(5), 7691-7709.
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5

Sorting Upper Palate Stem Cells

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For FACS, upper palates were dissected 4 weeks post chase from K5tetH2BGFP mice and labeled with PE conjugated anti-α6 integrin (CD49f) (1:200; BD Pharmingen) and APC conjugated CD44 (1:200); BD Pharminogen) for 30min and sorted using the FACS Aria II cell sorter (BD, Bioscience) for H2BGFP+/CD44+/α6+and H2BGFP−/CD44+/α6+ populations. Cells were collected in RNA protect Cell Reagent (Qiagen) for RNA isolation.
Zhang H., Boddupally K., Kandyba E., Kobielak K., Chen Y., Zu S., Krishnan R., Sinha U, & Kobielak A. (2014). Defining the localization and molecular characteristic of minor salivary gland label retaining cells. Stem cells (Dayton, Ohio), 32(8), 2267-2277.
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6

Flow Cytometry Analysis of Stem Cell Markers

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FACS analysis and cell sorting were performed using FACS Calibur and FACS Aria machines (Becton Dickinson, Palo Alto, CA), respectively. FACS data were analyzed using FlowJo software (Tree Star, Ashland, OR). Antibodies to the following proteins were used: APC-conjugated CD44 (BD Bioscience, Cat. 559942, dilution 1/40), PE-conjugated CD133 (MACS; Miltenyi Biotech, Sunnyvale, CA, 130-080-081, dilution 1/40), CD34 (MACS; Miltenyi Biotech, Sunnyvale, CA, 30-081-002,dilution 1/40), CD44 (MACS; Miltenyi Biotech, Sunnyvale, CA, 130-095-180, dilution 1/40), CD45 (MACS; Miltenyi Biotech, Sunnyvale, CA, 130-080-201, dilution 1/40), CD73 (MACS; Miltenyi Biotech, Sunnyvale, CA, 130-095-182, dilution 1/40), CD105 (MACS; Miltenyi Biotech, Sunnyvale, CA, 130-094-941, dilution 1/40), Annexin V (BD Bioscience, Cat. No. 556547, dilution 1/40), and propidium iodide (PI) (Invitrogen, Cat No P3566, dilution 1/40). Add 5 μl of antibody from each dilution into separate sample tubes containing cells (4 × 105). Mix well and incubate cells on ice for 30 min. Wash with 10 ml of medium. The FACS gates were established by staining with an isotype antibody.
Lee N.H., Cho A., Park S.R., Lee J.W., Sung Taek P., Park C.H., Choi Y.H., Lim S., Baek M.K., Kim D.Y., Jin M., Lee H.Y, & Hong I.S. (2018). SERPINB2 is a novel indicator of stem cell toxicity. Cell Death & Disease, 9(7), 724.
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7

Multiparametric Immunophenotyping of Stem Cells

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FACS analysis and cell sorting were performed using FACS Calibur and FACS Aria machines (Becton Dickinson, Palo Alto, CA), respectively. FACS data were analyzed using FlowJo software (Tree Star, Ashland, OR). Antibodies against the following proteins were used: APC-conjugated CD44 (BD Bioscience, Cat. 559942, dilution 1/40), PE-conjugated CD133 (MACS; Miltenyi Biotech, 130-080-081, dilution 1/40), CD34 (MACS; Miltenyi Biotech, 30-081-002), CD44 (MACS; Miltenyi Biotech, 130-095-180), CD45 (MACS; Miltenyi Biotech, 130-080-201), CD73 (MACS; Miltenyi Biotech, 130-095-182), CD105 (MACS; Miltenyi Biotech, 130-094-941), and CD140b (MACS; Miltenyi Biotech, 130-105-279). The FACS gates were established by staining with an isotype antibody or secondary antibody.
Park S.R., Kim S.K., Kim S.R., Kim D., Kim K.W., Hong I.S, & Lee H.Y. (2021). Noncanonical functions of glucocorticoids: A novel role for glucocorticoids in performing multiple beneficial functions in endometrial stem cells. Cell Death & Disease, 12(6), 612.
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8

Characterizing Stem Cell Markers

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To access the levels of Aldefluor, CD24, CD44 and CD133 positive populations, the cells were stained with Aldefluor kit (Stem Cell Technologies, Vancouver, Canada, Cat. 01700, dilution 1/40), PE conjugated CD24 (BD Bioscience, San. Jose, CA, USA, Cat. 555428, dilution 1/40), APC conjugated CD44 (BD Bioscience, Cat. 559942, dilution 1/40) and PE conjugated CD133 (MACS; Miltenyi Biotech, Sunnyvale, CA, USA, 130-080-081, dilution 1/40), respectively. Flow cytometry data were analyzed using FlowJo software (Tree Star, Ashland, OR, USA). The flow cytometry gates were established by staining with an isotype antibody or secondary antibody.
Lee N.H., Park S.R., Lee J.W., Lim S., Lee S.H., Nam S., Kim D.Y., Hah S.Y., Hong I.S, & Lee H.Y. (2019). SERPINB2 Is a Novel Indicator of Cancer Stem Cell Tumorigenicity in Multiple Cancer Types. Cancers, 11(4), 499.
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9

Multiparameter Flow Cytometry Profiling

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Freshly dissociated cells were stained with APC-conjugated EpCAM (dilution 1:100, Miltenyi clone HEA-125) and PerCP-Cy5.5-conjugated CD49f (dilution 1:10, BD clone GoH3); or APC-conjugated CD44 (dilution 1:10, BD clone G44-CD26) and PerCP-Cy5.5-conjugated CD24 (dilution 1:10, BD clone ML5); with live/dead Violet (dilution 1:1000, ThermoFisher) for cell viability, in HBSS (Gibco) with 2% FBS and incubated at room temperature for 20 min, followed by washing in HBSS with 2% FBS and re-suspended in HBSS/FBS 2%. Analysis was performed by using a CyAn (Beckman Coulter) flow cytometer. Thresholds on fluorescence signal intensity (subtracting background fluorescence from the appropriate isotype control antibodies) were used to determine the proportion of cell populations. Data were analyzed with FlowJo software.
Laisné M., Rodgers B., Benlamara S., Wicinski J., Nicolas A., Djerroudi L., Gupta N., Ferry L., Kirsh O., Daher D., Philippe C., Okada Y., Charafe-Jauffret E., Cristofari G., Meseure D., Vincent-Salomon A., Ginestier C, & Defossez P.A. (2024). A novel bioinformatic approach reveals cooperation between Cancer/Testis genes in basal-like breast tumors. Oncogene, 43(18), 1369-1385.
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