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Clone hir2

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Clone HIR2 is a monoclonal antibody that recognizes the human transferrin receptor (CD71). It is suitable for use in flow cytometry, immunohistochemistry, and other immunoassays to detect and quantify cells expressing CD71.

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Lab products found in correlation

C kit apccy7, by BioLegend (2 mentions) Cd14 fitc, by BioLegend (2 mentions) Cd45 pe, by BioLegend (2 mentions) Cd45.1 apc, by BioLegend (2 mentions) Cd15 pacific blue, by BioLegend (2 mentions) Glycophorin pecy5, by BioLegend (2 mentions) Mac 1 cd11b pecy7, by BioLegend (2 mentions) Gr 1 pe, by BioLegend (2 mentions) B220 pecy5, by BioLegend (2 mentions) Cd34 apc, by BioLegend (2 mentions) Cd45.2 fitc, by BioLegend (2 mentions) Facsaria 2, by BD (2 mentions) Sca1 apc, by BioLegend (2 mentions) Cd11b, by BioLegend (2 mentions) Clone hi30, by BioLegend (1 mentions) Streptavidin af488, by Thermo Fisher Scientific (1 mentions) Cd45 bv510, by BioLegend (1 mentions) Cd31 apc cy7, by BioLegend (1 mentions) Anti biotin magnetic beads, by Miltenyi Biotec (1 mentions) Cd235a pe cy7, by BioLegend (1 mentions)

3 protocols using clone hir2

1

Hematopoietic Progenitor Cell Analysis

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Cells were suspended in Ca++ Mg++-free phosphate-buffered saline (PBS) containing 2% (vol/vol) FBS (Globalstem). Human hematopoietic progenitors cells were identified by labeling with CD45-PE (1:50; clone HI30) and or CD34-APC (1:50; clone 561) while mature phenotypes were analyzed using CD15-Pacific Blue (monocytes and granulocytes) (1:50; clone W6D3/HI98), CD14-FITC (monocytes and macrophages)(clone 1:50; M5E2); CD11b (1:50; ICRF44) (macrophages) and Glycophorin-PECy5 (red blood cell membrane), and erythroid precursors (1:50; Clone HIR2) (all from Biolegend). Murine hematopoietic cells were identified using the following antibodies: Mac-1/CD11b-PECy7 (1:200; clone M1/70), Gr-1-PE (1:200; clone 8C5), B220-PECy5 (1:100; clone RA3-6B2), Sca1-APC (1:100; clone E13-161-7), c-Kit-APCCy7 (1:100; clone 2B8): CD45.1-APC (1:100; clone A20) and CD45.2-FITC (1:100; clone 104) (all from Biolegends). All antibodies were incubated at the concentration suggested by the manufacturer for 30 minutes on ice. Non-specific signals and dead cells were excluded by appropriate fluorochrome-conjugated isotype and propidium iodide staining, respectively. Cell fluorescence was analyzed with the FACSAria II (Becton Dickinson).
Amabile G., Di Ruscio A., Müller F., Welner R.S., Yang H., Ebralidze A.K., Zhang H., Levantini E., Qi L., Martinelli G., Brummelkamp T., Le Beau M.M., Figueroa M.E., Bock C, & Tenen D.G. (2015). Dissecting the role of aberrant DNA methylation in human leukemia. Nature communications, 6, 7091.
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2

Hematopoietic Progenitor Cell Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were suspended in Ca++ Mg++-free phosphate-buffered saline (PBS) containing 2% (vol/vol) FBS (Globalstem). Human hematopoietic progenitors cells were identified by labeling with CD45-PE (1:50; clone HI30) and or CD34-APC (1:50; clone 561) while mature phenotypes were analyzed using CD15-Pacific Blue (monocytes and granulocytes) (1:50; clone W6D3/HI98), CD14-FITC (monocytes and macrophages)(clone 1:50; M5E2); CD11b (1:50; ICRF44) (macrophages) and Glycophorin-PECy5 (red blood cell membrane), and erythroid precursors (1:50; Clone HIR2) (all from Biolegend). Murine hematopoietic cells were identified using the following antibodies: Mac-1/CD11b-PECy7 (1:200; clone M1/70), Gr-1-PE (1:200; clone 8C5), B220-PECy5 (1:100; clone RA3-6B2), Sca1-APC (1:100; clone E13-161-7), c-Kit-APCCy7 (1:100; clone 2B8): CD45.1-APC (1:100; clone A20) and CD45.2-FITC (1:100; clone 104) (all from Biolegends). All antibodies were incubated at the concentration suggested by the manufacturer for 30 minutes on ice. Non-specific signals and dead cells were excluded by appropriate fluorochrome-conjugated isotype and propidium iodide staining, respectively. Cell fluorescence was analyzed with the FACSAria II (Becton Dickinson).
Amabile G., Di Ruscio A., Müller F., Welner R.S., Yang H., Ebralidze A.K., Zhang H., Levantini E., Qi L., Martinelli G., Brummelkamp T., Le Beau M.M., Figueroa M.E., Bock C, & Tenen D.G. (2015). Dissecting the role of aberrant DNA methylation in human leukemia. Nature communications, 6, 7091.
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3

Immunophenotyping of Hematopoietic Subsets

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Frozen BM aspirates were thawed on ice and stained with biotinylated antibodies against CD45 (1:50, clone HI30; BioLegend) and CD235a (1:50, clone HIR2, BioLegend) followed by depletion using magnetic anti-biotin beads (20 μL per 107 cells; Miltenyi Biotec) in PBS supplemented with 2% FCS and iMag (BD Biosciences). After depletion, the remaining cells were stained in PBS containing 0.5% FCS at 4°C with the following antibodies: Streptavidin-AF488 (1:100, Invitrogen), CD45-BV510(1:50, clone HI30; BioLegend), CD235a-PE-Cy7(1:50, clone HI264; BioLegend), CD71-AF700 (1:20, clone MEM-75; Exbio), CD271-PE (1:50, clone ME20.4; BioLegend), CD31-APC-Cy7 (1:20, clone WM59; BioLegend), and CD44-APC (1:50, clone IM7, Sony). 7AAD (1:100; Beckman Coulter) was used for dead cell exclusion. Samples were measured using FACSymphony A5 Cell Analyzer and analyzed with a FlowJo_v10.6.1 program.
Chen L., Pronk E., van Dijk C., Bian Y., Feyen J., van Tienhoven T., Yildirim M., Pisterzi P., de Jong M.M., Bastidas A., Hoogenboezem R.M., Wevers C., Bindels E.M., Löwenberg B., Cupedo T., Sanders M.A, & Raaijmakers M.H. (2023). A Single-Cell Taxonomy Predicts Inflammatory Niche Remodeling to Drive Tissue Failure and Outcome in Human AML. Blood Cancer Discovery, 4(5), 394-417.
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