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4 protocols using cd47 pe

1

Multimarker Profiling of Cell Lineages

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The primary anti-human or anti-mouse antibodies (CD29-PE, CD49a-PE, CD49b-PE, CD49c-PE, CD49d-FITC, CD49e-PE, CD49f-PE, CD51/61-PE, CD44-APC, CD45-FITC, CD47-PE, CD324-PE, CD325-PE, CD326-PE, CD54-PE, CD106-FITC, CD62E-APC, CD62L-APC, CD15s, isotype controls) and secondary antibodies were all obtained from Becton Dickinson (BD) Pharmingen™. Human interleukin-1 beta (IL-1β) was purchased from Cell Signaling Technology, Inc. 17β-estradiol (E2) and progesterone (P4) were purchased from Shanghai Yuanye biological technology Co., Ltd Mifepristone (RU-486) was purchased from Shanghai New Hualian pharmaceutical Co., Ltd with purity > 98%. Metapristone was synthesized by our laboratory with purity > 98%. Calcein-AM was obtained from Dojindo Molecular Technologies, Inc.
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2

Flow Cytometric Analysis of Cell Surface Markers

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The primary anti-human or anti-mouse antibodies (CD29-PE, CD49a-PE, CD49b-PE, CD49c-PE, CD49d-FITC, CD49e-PE, CD49f-PE, CD51/61-PE, CD44-APC, CD45-FITC, CD47-PE, CD324-PE, CD325-PE, CD326-PE, CD54-PE, CD106-FITC, CD62E-APC, CD62L-APC, CD15s, isotype controls) and secondary antibodies were all obtained from Becton Dickinson (BD) Pharmingen™. Human interleukin-1 beta (IL-1β) was purchased from Cell Signaling Technology, Inc. 17β-estradiol (E2) and progesterone (P4) were purchased from Shanghai Yuanye biological technology Co. Mifepristone (RU-486) was purchased from Shanghai New Hualian pharmaceutical Co., with purity > 98%. Metapristone was synthesized by our laboratory with purity > 98%. Calcein-AM was obtained from Dojindo Molecular Technologies, Inc.
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3

Flow Cytometry of Tumor Cell Markers

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Cell surface molecules on tumor cell lines and human macrophages were measured by flow cytometry using the following fluorochrome-conjugated antibodies: CD47-PE, CD47-APC, PD-L1-PE, and CD11b-PE (BD Biosciences, Franklin Lakes, NJ, USA). CD172a-PerCP-eF710 was purchased from eBioscience (Thermo Fisher Scientific, Waltham, MA). Flow cytometry analysis was performed using a FACS Calibur or FACS Canto II (BD Biosciences), and the data were analyzed using FlowJo v10.7 software (FlowJo LLC, Ashland, OR, USA, https://www.flowjo.com/).
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4

Flow Cytometry Characterization of Leukocytes

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Antibodies from BD Biosciences; CD15-BV650 (#563142, clone W6D3), CD45-PE-CF594 (#562279 clone HI30), CD16-V450 (#560474 clone 3G8), CD62L-BV510 (#563203 clone DREG-56), CD47-PE (#556046 clone B6H12), CD36-APC (#550956 clone CB38), CD11b-APC-Cy7 (#557754 clone ICRF44), CD66b-PerCP-Cy5.5 (#562254 clone G10F5), CD49d-BV711(#563177 clone 9F10), CD43-BB515 (#564542 clone 1G10) and CD24-PE-Cy7 (#561646 clone ML5).
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