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Cd34 pe clone 563

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CD34-PE (clone 563) is a laboratory reagent used in flow cytometry applications. It is a fluorescently-labeled antibody that binds to the CD34 antigen, which is expressed on hematopoietic stem and progenitor cells. This product can be used to identify and quantify CD34-positive cells in various biological samples.

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Lab products found in correlation

Cd45 apc clone hi30, by BD (7 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (7 mentions) Mcd45 pe cy7 clone 30 f11, by BD (4 mentions) Fortessa, by BD (4 mentions) Cd19 pe clone hib19, by BD (3 mentions) Cd33 bv421 clone wm53, by BD (3 mentions) Facsaria 2, by BD (3 mentions) Cd15 bv785 clone w6d3, by BioLegend (2 mentions) Cd38 pe cf594 clone hit2, by BD (2 mentions) Cd41a pe cy7 clone hip8, by BD (1 mentions) Cd33 pe cf594 clone wm53, by BD (1 mentions) Cytofix cytoperm plus kit, by BD (1 mentions) Cd38 percpcy5.5 clone hit2, by BD (1 mentions) Igd pe clone ia6 2, by BD (1 mentions) Human fc block, by BD (1 mentions) 7 aad viability staining solution, by Thermo Fisher Scientific (1 mentions) Cd86 pe clone it2, by BD (1 mentions) Cd69 pe cy7 clone fn50, by BD (1 mentions) Facsverse flow cytometer, by BD (1 mentions) Cd19 apc clone hib19, by BioLegend (1 mentions)

8 protocols using cd34 pe clone 563

1

Comprehensive Immune Cell Profiling

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The following antibodies were used: CD34-PE (clone 563, BD Pharmingen), CD45-APC (clone HI30, BD PharMingen), CD33-BV421 (clone WM53, BD Horizon), mCD45-PE-Cy7 (clone 30-F11, BD Biosciences), CD19-PE (clone HIB19, BD Pharmingen), CD38-PE-CF594 (clone HIT2, BD Horizon), CD15-BV785 (clone W6D3, BioLegend), CD14-APC (clone M5E2, BD Horizon), and CD44-APC. Cell viability was assessed with DAPI (Life Technologies). Cells were assayed on a BD Fortessa and data were analyzed with FlowJo software (Tree Star). Cell sorting was performed on a BD FACSAria II.
Kotini A.G., Carcamo S., Cruz-Rodriguez N., Olszewska M., Wang T., Demircioglu D., Chang C.J., Bernard E., Chao M.P., Majeti R., Luo H., Kharas M.G., Hasson D, & Papapetrou E.P. (2023). Patient-Derived iPSCs Faithfully Represent the Genetic Diversity and Cellular Architecture of Human Acute Myeloid Leukemia. Blood Cancer Discovery, 4(4), 318-335.
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2

Multicolor Flow Cytometry Analysis

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The antibodies CD34-PE (clone 563, BD Pharmingen) and CD45-APC (clone HI30, BD PharMingen) were used. Cell viability was assessed with DAPI (Life Technologies). Cells were then assayed on a BD Fortessa, and data were analyzed with FlowJo software (Tree Star).
Kotini A.G, & Papapetrou E.P. (2020). Engineering of targeted megabase-scale deletions in human induced pluripotent stem cells. Experimental hematology, 87, 25-32.
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3

Flow Cytometry Analysis of Hematopoietic Cells

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Following hematopoietic differentiation, the cells were dissociated with accutase into single cells. The following flow cytometry antibodies were used: CD34-PE (clone 563, BD PharMingen), CD45-BV510 (clone HI30, BD Horizon), CD90-PerCP/Cy5.5 (clone 5E10, BioLegend), CD38-PE-CF594 (clone HIT2, BD Horizon), CD33-PE-CF594 (clone WM53, BD Horizon), CD49f-BV421 (clone GoH3, BD Horizon), CD45RA-APC (clone MEM56, Life Technologies), CD123-BV711 (clone 9F5, BD Horizon), CD41a- PE-Cy7 (clone HIP8, BD PharMingen), mCD45-PE-Cy7 (clone 30-F11, BD Biosciences), CD45-APC (clone HI30, BD PharMingen), CD19-PerCP-Cy5.5 (clone HIB19, BD PharMingen). Cell viability was assessed with DAPI (Life Technologies). Intracellular staining of Patient 65 cells was performed with a biotin-conjugated TSPAN18 antibody (Lifespan Bioscience) and a secondary ani-biotin APC antibody (Miltenyi Biotec) using the Cytofix/Cytoperm Plus kit (BD Biosciences). Cells were assayed on a BD Fortessa and data were analyzed with FlowJo software (Tree Star).
Wesely J., Kotini A.G., Izzo F., Luo H., Yuan H., Sun J., Georgomanoli M., Zviran A., Deslauriers A.G., Dusaj N., Nimer S.D., Leslie C., Landau D.A., Kharas M.G, & Papapetrou E.P. (2020). Acute Myeloid Leukemia iPSCs Reveal a Role for RUNX1 in the Maintenance of Human Leukemia Stem Cells. Cell reports, 31(9), 107688.
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4

Multiparametric Flow Cytometry Analysis

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Single-cell suspensions from the BM and peripheral blood were first incubated with human BD Fc block (BD Biosciences) to block Fcγ receptor and then stained with the following anti-human Abs: CD19-APC (clone HIB19, BioLegend), CD34-PE (clone 563, BD Biosciences), CD20-FITC (clone 2H7, BioLegend), CD27-APC/Cyanine7 (clone O323, BioLegend), IgD-PE (clone IA6-2, BD Biosciences), IgM-FITC (clone G20-127, BD Biosciences), CD43-PE (clone CD43-10G7, BioLegend), CD38-PE/Cy7 (clone HIT2, BioLegend), CD38-Percp/Cy5.5 (clone HIT2, BD Biosciences), CD24-PE (clone ML5, BD Biosciences), CD27-V450 (clone M-T271, BD Biosciences), IgD-BV510 (clone IA6-2, BD Biosciences), IgG-PE/Cy7 (clone G18-145, BD Biosciences), Ki-67–PE (clone Ki-67, BioLegend), CD80-FITC (clone L307.4, BD Biosciences), CD86-PE (clone IT2.2, BD Biosciences), HLADR-PE (clone G46-6, BD Biosciences) and CD69-PE/Cy7 (clone FN50, BD Biosciences). 7-AAD Viability Staining Solution (eBioscience, Thermo Fisher Scientific) was used for live versus dead cell discrimination. Samples were analyzed with a FACSVerse flow cytometer (BD Biosciences) using the FACSuite software. Data analysis was performed with FlowJo 10 software (Treestar).
Min Q., Meng X., Zhou Q., Wang Y., Li Y., Lai N., Xiong E., Wang W., Yasuda S., Yu M., Zhang H., Sun J., Wang X, & Wang J.Y. (2021). RAG1 splicing mutation causes enhanced B cell differentiation and autoantibody production. JCI Insight, 6(19), e148887.
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5

Flow Cytometry Immunophenotyping Protocol

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The following antibodies were used: CD34-PE (clone 563, BD Pharmingen, RRID:AB_393871), CD45-APC (clone HI30, BD Pharmingen, RRID:AB_398600), CD14-APC (clone M5E2, BD Pharmingen, RRID:AB_398596), CD15-BV785 (clone W6D3, BioLegend, RRID:AB_2632921), CD16-BV510 (clone 3G8, BD Horizon, RRID:AB_2744296). Cell viability was assessed with DAPI (Life Technologies). Cells were assayed on a BD Fortessa and data were analyzed with FlowJo software (Tree Star, RRID:SCR_008520).
Wheeler E.C., Vora S., Mayer D., Kotini A.G., Olszewska M., Park S.S., Guccione E., Teruya-Feldstein J., Silverman L., Sunahara R.K., Yeo G.W, & Papapetrou E.P. (2022). Integrative RNA-omics discovers GNAS alternative splicing as a phenotypic driver of splicing factor-mutant neoplasms. Cancer discovery, 12(3), 836-855.
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6

Immunophenotyping of Hematopoietic Cells

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The following antibodies were used: CD34-PE (clone 563, BD PharMingen), CD45-APC (clone HI30, BD PharMingen), CD33-BV421 (clone WM53, BD Horizon), mCD45-PE-Cy7 (clone 30-F11, BD Biosciences), CD19-PE (clone HIB19, BD PharMingen). Cell viability was assessed with DAPI (Life Technologies). Cells were assayed on a BD Fortessa and data were analyzed with FlowJo software (Tree Star). Cell sorting was performed on a BD FACS Aria II.
Wang T., Pine A.R., Kotini A.G., Yuan H., Zamparo L., Starczynowski D.T., Leslie C, & Papapetrou E.P. (2021). Sequential CRISPR gene editing in human iPSCs charts the clonal evolution of myeloid leukemia and identifies early disease targets. Cell stem cell, 28(6), 1074-1089.e7.
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7

Multicolor Flow Cytometry Analysis

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The antibodies CD34-PE (clone 563, BD Pharmingen) and CD45-APC (clone HI30, BD PharMingen) were used. Cell viability was assessed with DAPI (Life Technologies). Cells were then assayed on a BD Fortessa, and data were analyzed with FlowJo software (Tree Star).
Kotini A.G, & Papapetrou E.P. (2020). Engineering of targeted megabase-scale deletions in human induced pluripotent stem cells. Experimental hematology, 87, 25-32.
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8

iPSC-Derived HSPC Transplantation in NSG Mice

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The following antibodies were used: CD34-PE (clone 563, BD Pharmingen), CD45-APC (clone HI30, BD PharMingen), CD33-BV421 (clone WM53, BD Horizon), mCD45-PE-Cy7 (clone 30-F11, BD Biosciences), CD19-PE (clone HIB19, BD Pharmingen). Cell viability was assessed with DAPI (Life Technologies). Cells were assayed on a BD Fortessa and data were analyzed with FlowJo software (Tree Star). Cell sorting was performed on a BD FACS Aria II.
Transplantation into NSG mice NSG (NOD.Cg-Prkdc scid Il2rg tm1Wjl /SzJ) and NSG-SGM3 (NOD.Cg-Prkdc scid Il2rg tm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/MloySzJ) mice were purchased from Jackson Laboratories and housed at the Center for Comparative Medicine and Surgery at Icahn School of Medicine at Mount Sinai. One day before transplantation, the mice were injected intraperitoneally with 30mg/kg Busulfan solution. P, A, SA, SAR and SARF iPSC-HSPCs from day 13-15 of hematopoietic differentiation were resuspended in StemPro-34 and injected via the tail vein using a 25G needle at 1x10 6 cells per 100µL per mouse. The mice were sacrificed after 13-15 weeks. Bone marrow was collected from the femurs and tibia for flow cytometry and spleens were harvested and their weight recorded. All mouse studies were performed in compliance with Icahn School of Medicine at Mount Sinai laboratory animal care regulations.
Wang T., Pine A.R., Wesely J., Yuan H., Zamparo L., Kotini A.G., Leslie C., & Papapetrou E.P. (2020). Sequential CRISPR gene editing in human iPSCs charts the clonal evolution of leukemia.
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