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Cd34 apc

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The CD34-APC is a flow cytometry antibody reagent that detects the CD34 antigen, a marker for hematopoietic stem and progenitor cells. It is conjugated to the fluorescent dye allophycocyanin (APC) for detection in flow cytometry applications.

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Lab products found in correlation

Facscalibur, by BD (7 mentions) Anti cd71 fitc, by Beckman Coulter (7 mentions) Anti cd41 fitc gpiib iiia, by BD (7 mentions) Anti cd33 fitc, by Beckman Coulter (7 mentions) Anti cd41 pe, by Beckman Coulter (7 mentions) Anti cd45 percp, by Beckman Coulter (7 mentions) Trypsin edta, by Thermo Fisher Scientific (1 mentions) Vegfr 2 pe, by R&D Systems (1 mentions) Egm 2 medium, by Lonza (1 mentions) 7 aad, by Beckman Coulter (1 mentions) Fc500 flow cytometer, by Beckman Coulter (1 mentions) Fitc cd38, by Beckman Coulter (1 mentions) Cd10 fitc, by Beckman Coulter (1 mentions) Facsaria sorp flow cytometer, by BD (1 mentions) Diva software, by BD (1 mentions) Repli g single cell kit, by Qiagen (1 mentions) Cd45ra pb, by BioLegend (1 mentions) Cd38 pe cy7, by BioLegend (1 mentions) Cd123 pe, by BioLegend (1 mentions)

10 protocols using cd34 apc

1

Multiparametric Analysis of Platelet Differentiation

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Example 34

The differentiation state of platelets in culture can be assessed by flow cytometry. Megakaryocytes (MKs) represent a distinct cellular morphology that precedes terminal platelet differentiation. To determine the extent of maturation toward MKs, 1×10^6 cultured cells (LAMA-84 and CD34+ cells) are washed and then labeled with (a) anti-CD41-FITC (GpIIb/IIIa; BD Bioscience, San Jose, Calif., USA) or anti CD71-FITC or (b) anti-CD33-FITC, anti-CD41-PE, anti-CD45-PerCp and CD34-APC (Beckman Coulter, Fullerton, Calif., USA), and analyzed for the percentage of CD41 cells generated.

To determine the amount of ploidy, differentiated LAMA-84 cells are fixed overnight in 75% ethanol at 4° C. and labeled with propidium iodide (PI, 50 μg/ml) and analyzed using the FACScalibur (Becton Dickinson), whereas day 14 differentiated CD34+ cells are analyzed quantitatively under a microscope after May-Grunwald/Giemsa staining by quantitating the number of nuclei per cell and specific morphology of MKs with this stain. Only cells with MK morphology are analyzed. The presence of multinucleated cells in the cytospin preparation is indicative of the presence of polyploid MKs. Differentiated CD34+ cells are assessed for the presence of multinucleated mature MKs by morphology.

US10253296B2. Synthetic membrane-receiver complexes (2019-04-09). RUBIUS THERAPEUTICS, INC. [US]. Inventors: Avak Kahvejian [US], Jordi Mata-Fink [US], John Round [US], David Arthur Berry [US], Noubar B. Afeyan [US].
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2

Characterizing Endothelial CD109 Expression

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To validate by flow-cytometry the expression of CD109 on endothelial cells, in vitro endothelial colonies were obtained from peripheral blood (PB) of 10 healthy donors (7 female, 3 male) and 23 cancer patients as previously described [32] (link).
Briefly, mononuclear cells (MNCs) were isolated from PB using Ficoll-Paque gradient centrifugation, resuspended into EGM2 medium (Lonza, Walkersville, MD) and seeded onto Collagen I petri dishes (35 mm, Biocoat, BD Labware, Bedford, MA). Cultures were incubated at 37°C, 5% CO2, 95% relative humidity for 3–4 weeks. Medium was changed every 2 days for 7 days and then twice a week until first passage, and culture monitored for the detection of endothelial colonies on the basis of morphological features, as previously described [32] (link). After 15–20 days, cells were detached with trypsin/EDTA (Gibco, BRL, UK) and stained with monoclonal antibodies (MoAbs) anti- CD31-PeCy7, CD34-APC, CD45-H7, and 7-AAD (all from Beckman Coulter) CD146-Pe (Chemicon), VEGFR-2- PE (R&D) and CD109-Pe (Abcam) for flow cytometry studies.
Mancuso P., Calleri A., Gregato G., Labanca V., Quarna J., Antoniotti P., Cuppini L., Finocchiaro G., Eoli M., Rosti V, & Bertolini F. (2014). A Subpopulation of Circulating Endothelial Cells Express CD109 and is Enriched in the Blood of Cancer Patients. PLoS ONE, 9(12), e114713.
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3

Quantifying Leukemia Stem Cell Subpopulations

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To determine the effect on LSCs, primary AML, ALL, or CLL cells were cultured in the presence or absence of test compound, washed with 1 × PBS and stained with CD34−APC and either CD38−FITC or CD10−FITC (Beckman Coulter, Brea, CA) on ice for 1 hour. All antibodies were used at a final concentration of 200 ng/mL. Stained cells were analyzed on a Beckman Coulter FC500 flow cytometer. Phenotypically described stem cell subpopulations were quantified by gating the CD34+CD38− population for AML24 (link) and CLL25 (link) and the CD34+CD10− population for ALL.6 (link) To determine human cell engraftment and the effect of test compound on stem cell populations in vivo, mice were euthanized 8 weeks post transplantation. Bone marrow (BM) cells were isolated by flushing femurs and tibias of the mice with 1 × PBS. BM cells were stained with antibodies against HLA (200 ng/mL, BD Biosciences, San Jose, CA), CD34, CD38 and CD10.
Gunasekara D.C., Zheng M.M., Mojtahed T., Woods J.R., Fandy T.E., Riofski M.V., Glackin C.A., Hassan H.E., Kirshner J, & Colby D.A. (2016). 15-Methylene-Eburnamonine Kills Leukemic Stem Cells and Reduces Engraftment in a Humanized Bone Marrow Xenograft Mouse Model of Leukemia. ChemMedChem, 11(21), 2392-2397.
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4

Assessing Platelet Differentiation by Flow Cytometry

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Example 44

The differentiation state of platelets in culture can be assessed by flow cytometry. Megakaryocytes (MKs) represent a distinct cellular morphology that precedes terminal platelet differentiation. To determine the extent of maturation toward MKs, 1×10{circumflex over ( )}6 cultured cells (LAMA-84 and CD34+ cells) are washed and then labeled with (a) anti-CD41-FITC (GpIIb/IIIa; BD Bioscience, San Jose, Calif., USA) or anti CD71-FITC or (b) anti-CD33-FITC, anti-CD41-PE, anti-CD45-PerCp and CD34-APC (Beckman Coulter, Fullerton, Calif., USA), and analyzed for the percentage of CD41 cells generated.

To determine the amount of ploidy, differentiated LAMA-84 cells are fixed overnight in 75% ethanol at 4° C. and labeled with propidium iodide (PI, 50 ug/ml) and analyzed using the FACScalibur (Becton Dickinson), whereas day 14 differentiated CD34+ cells are analyzed quantitatively under a microscope after May-Grunwald/Giemsa staining by quantitating the number of nuclei per cell and specific morphology of MKs with this stain. Only cells with MK morphology are analyzed. The presence of multinucleated cells in the cytospin preparation is indicative of the presence of polyploid MKs. Differentiated CD34+ cells are assessed for the presence of multinucleated mature MKs by morphology.

US11554141B2. Methods and compositions for immunomodulation (2023-01-17). RUBIUS THERAPEUTICS, INC. [US]. Inventors: Jordi Mata-Fink [US], John Round [US], Noubar B. Afeyan [US], Avak Kahvejian [US].
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5

Assessing Platelet Differentiation by Flow Cytometry

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Example 34

The differentiation state of platelets in culture can be assessed by flow cytometry. Megakaryocytes (MKs) represent a distinct cellular morphology that precedes terminal platelet differentiation. To determine the extent of maturation toward MKs, 1×10{circumflex over ( )}6 cultured cells (LAMA-84 and CD34+ cells) are washed and then labeled with (a) anti-CD41-FITC (GpIIb/IIIa; BD Bioscience, San Jose, Calif., USA) or anti CD71-FITC or (b) anti-CD33-FITC, anti-CD41-PE, anti-CD45-PerCp and CD34-APC (Beckman Coulter, Fullerton, Calif., USA), and analyzed for the percentage of CD41 cells generated.

To determine the amount of ploidy, differentiated LAMA-84 cells are fixed overnight in 75% ethanol at 4° C. and labeled with propidium iodide (PI, 50 μg/ml) and analyzed using the FACScalibur (Becton Dickinson), whereas day 14 differentiated CD34+ cells are analyzed quantitatively under a microscope after May-Grunwald/Giemsa staining by quantitating the number of nuclei per cell and specific morphology of MKs with this stain. Only cells with MK morphology are analyzed. The presence of multinucleated cells in the cytospin preparation is indicative of the presence of polyploid MKs. Differentiated CD34+ cells are assessed for the presence of multinucleated mature MKs by morphology.

US10557119B2. Erythroid cells comprising phenylalanine ammonia lyase (2020-02-11). RUBIUS THERAPEUTICS, INC. [US]. Inventors: Avak Kahvejian [US], Jordi Mata-Fink [US], John Round [US], David Arthur Berry [US], Noubar B. Afeyan [US].
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6

Assessing Platelet Differentiation by Flow Cytometry

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Example 34

The differentiation state of platelets in culture can be assessed by flow cytometry. Megakaryocytes (MKs) represent a distinct cellular morphology that precedes terminal platelet differentiation. To determine the extent of maturation toward MKs, 1×10{circumflex over ( )}6 cultured cells (LAMA-84 and CD34+ cells) are washed and then labeled with (a) anti-CD41-FITC (GpIIb/IIIa; BD Bioscience, San Jose, Calif., USA) or anti CD71-FITC or (b) anti-CD33-FITC, anti-CD41-PE, anti-CD45-PerCp and CD34-APC (Beckman Coulter, Fullerton, Calif., USA), and analyzed for the percentage of CD41 cells generated.

To determine the amount of ploidy, differentiated LAMA-84 cells are fixed overnight in 75% ethanol at 4° C. and labeled with propidium iodide (PI, 50 μg/ml) and analyzed using the FACScalibur (Becton Dickinson), whereas day 14 differentiated CD34+ cells are analyzed quantitatively under a microscope after May-Grunwald/Giemsa staining by quantitating the number of nuclei per cell and specific morphology of MKs with this stain. Only cells with MK morphology are analyzed. The presence of multinucleated cells in the cytospin preparation is indicative of the presence of polyploid MKs. Differentiated CD34+ cells are assessed for the presence of multinucleated mature MKs by morphology.

US10344263B2. Synthetic membrane-receiver complexes (2019-07-09). RUBIUS THERAPEUTICS, INC. [US]. Inventors: Avak Kahvejian [US], Jordi Mata-Fink [US], John Round [US], David Arthur Berry [US], Noubar B. Afeyan [US].
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7

Multiparametric Analysis of Platelet Differentiation

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Example 34

The differentiation state of platelets in culture can be assessed by flow cytometry. Megakaryocytes (MKs) represent a distinct cellular morphology that precedes terminal platelet differentiation. To determine the extent of maturation toward MKs, 1×10^6 cultured cells (LAMA-84 and CD34+ cells) are washed and then labeled with (a) anti-CD41-FITC (GpIIb/IIIa; BD Bioscience, San Jose, Calif., USA) or anti CD71-FITC or (b) anti-CD33-FITC, anti-CD41-PE, anti-CD45-PerCp and CD34-APC (Beckman Coulter, Fullerton, Calif., USA), and analyzed for the percentage of CD41 cells generated.

To determine the amount of ploidy, differentiated LAMA-84 cells are fixed overnight in 75% ethanol at 4° C. and labeled with propidium iodide (PI, 50 μg/ml) and analyzed using the FACScalibur (Becton Dickinson), whereas day 14 differentiated CD34+ cells are analyzed quantitatively under a microscope after May-Grunwald/Giemsa staining by quantitating the number of nuclei per cell and specific morphology of MKs with this stain. Only cells with MK morphology are analyzed. The presence of multinucleated cells in the cytospin preparation is indicative of the presence of polyploid MKs. Differentiated CD34+ cells are assessed for the presence of multinucleated mature MKs by morphology.

US10301594B1. Synthetic membrane-receiver complexes (2019-05-28). RUBIUS THERAPEUTICS, INC. [US]. Inventors: Avak Kahvejian [US], Jordi Mata-Fink [US], John Round [US], David Arthur Berry [US], Noubar B. Afeyan [US].
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8

Myeloid Progenitor Cell Isolation

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1 ml viably frozen bone marrow MNCs were thawed in the presence of 100 μl DNAse I (2 mg μl−1) and incubated for 10 min in a solution of 1.6 ml fetal calf serum, 10 μl heparin (5,000 U ml−1) and 100 μl MgSO4 (0.22 μM). Subsequently, the myeloid progenitor cells were sorted according to a protocol adapted from Pang et al.35 (link) The cells were washed and stained with CD34-APC (Beckman Coulter, Brea, CA, USA), CD38-PE-Cy7 (BioLegend, San Diego, CA, USA), CD123-PE (BioLegend) and CD45RA-PB (BioLegend) monoclonal antibodies. Cells were analysed and sorted using a FACS Aria SORP flow cytometer and DIVA software (Becton Dickinson, Franklin Lakes, NJ, USA). Viable cells were selected based on forward scatter and side scatter profiles, and doublets were discriminated using forward scatter area versus width and side scatter area versus width. The HSC population was defined as CD34+CD38−. Within the CD34+CD38+ fraction, the common myeloid progenitor cells (CD123+CD45RA−), the granulocyte-macrophage progenitor cells (CD123+CD45RA+) and the megakaryocyte-erythroid progenitor cells (CD123−CD45RA−) were selected. DNA isolation from these cell fractions, followed by DNA amplification, was carried out using the Qiagen REPLI-g single cell kit (Qiagen) according to the manufacturer’s protocol.
da Silva-Coelho P., Kroeze L.I., Yoshida K., Koorenhof-Scheele T.N., Knops R., van de Locht L.T., de Graaf A.O., Massop M., Sandmann S., Dugas M., Stevens-Kroef M.J., Cermak J., Shiraishi Y., Chiba K., Tanaka H., Miyano S., de Witte T., Blijlevens N.M., Muus P., Huls G., van der Reijden B.A., Ogawa S, & Jansen J.H. (2017). Clonal evolution in myelodysplastic syndromes. Nature Communications, 8, 15099.
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9

Platelet Differentiation Analysis by Flow Cytometry

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Example 44

The differentiation state of platelets in culture can be assessed by flow cytometry. Megakaryocytes (MKs) represent a distinct cellular morphology that precedes terminal platelet differentiation. To determine the extent of maturation toward MKs, 1×10{circumflex over ( )}6 cultured cells (LAMA-84 and CD34+ cells) are washed and then labeled with (a) anti-CD41-FITC (GpIIb/IIIa; BD Bioscience, San Jose, Calif., USA) or anti CD71-FITC or (b) anti-CD33-FITC, anti-CD41-PE, anti-CD45-PerCp and CD34-APC (Beckman Coulter, Fullerton, Calif., USA), and analyzed for the percentage of CD41 cells generated.

To determine the amount of ploidy, differentiated LAMA-84 cells are fixed overnight in 75% ethanol at 4° C. and labeled with propidium iodide (PI, 50 μg/ml) and analyzed using the FACScalibur (Becton Dickinson), whereas day 14 differentiated CD34+ cells are analyzed quantitatively under a microscope after May-Grunwald/Giemsa staining by quantitating the number of nuclei per cell and specific morphology of MKs with this stain. Only cells with MK morphology are analyzed. The presence of multinucleated cells in the cytospin preparation is indicative of the presence of polyploid MKs. Differentiated CD34+ cells are assessed for the presence of multinucleated mature MKs by morphology.

US11576934B2. Methods and compositions for immunomodulation (2023-02-14). RUBIUS THERAPEUTICS, INC. [US]. Inventors: Jordi Mata-Fink [US], John Round [US], Noubar B. Afeyan [US], Avak Kahvejian [US].
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10

Assessing Platelet Differentiation and Maturation

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Example 34

The differentiation state of platelets in culture can be assessed by flow cytometry. Megakaryocytes (MKs) represent a distinct cellular morphology that precedes terminal platelet differentiation. To determine the extent of maturation toward MKs, 1×10^6 cultured cells (LAMA-84 and CD34+ cells) are washed and then labeled with (a) anti-CD41-FITC (GpIIb/IIIa; BD Bioscience, San Jose, Calif., USA) or anti CD71-FITC or (b) anti-CD33-FITC, anti-CD41-PE, anti-CD45-PerCp and CD34-APC (Beckman Coulter, Fullerton, Calif., USA), and analyzed for the percentage of CD41 cells generated.

To determine the amount of ploidy, differentiated LAMA-84 cells are fixed overnight in 75% ethanol at 4° C. and labeled with propidium iodide (PI, 50 μg/ml) and analyzed using the FACScalibur (Becton Dickinson), whereas day 14 differentiated CD34+ cells are analyzed quantitatively under a microscope after May-Grunwald/Giemsa staining by quantitating the number of nuclei per cell and specific morphology of MKs with this stain. Only cells with MK morphology are analyzed. The presence of multinucleated cells in the cytospin preparation is indicative of the presence of polyploid MKs. Differentiated CD34+ cells are assessed for the presence of multinucleated mature MKs by morphology.

US10329531B2. Synthetic membrane-receiver complexes (2019-06-25). RUBIUS THERAPEUTICS, INC. [US]. Inventors: Avak Kahvejian [US], Jordi Mata-Fink [US], John Round [US], David Arthur Berry [US], Noubar B. Afeyan [US].
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