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Click it flow cytometry assay kit

Manufactured by Thermo Fisher Scientific
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The Click-iT Flow Cytometry Assay kit is a tool used for the detection and quantification of cellular proliferation. It enables the measurement of DNA synthesis by labeling newly synthesized DNA with a clickable nucleoside analog.

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Lab products found in correlation

Op puro, by Jena Biosciences (3 mentions) Fortessa, by BD (2 mentions) Op puro, by BD (1 mentions) Lsr fortessa instrument, by BD (1 mentions) 5 ethynyl 2 deoxyuridine edu, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Click-iTTM Plus EdU Alexa FluorTM 647 flow cytometry assay kit Click-iTTM EdU Pacific BlueTM Flow Cytometry Assay Kit Click-iTTM Plus EdU flow Cytometry Assay Kit Click-iTTM EdU Flow Cytometry Assay Kit Click-iTTM EdU Alexa FluorTM 647 Flow Cytometry Assay Kit Flow cytometry

7 protocols using click it flow cytometry assay kit

1

Measuring Cellular Protein Synthesis

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Cells were plated at a density of 200,000 cells mL−1 and pre-treated with DMSO or Ro up to 4 h. Then, 50 μM O-propargyl-puromycin (OP-Puro; NU-931–05, Jena Bioscience) was added. Control cells were co-incubated with DMSO or Ro and treated with 150 μg mL-1 cycloheximide for 15 min. Non-OP-Puro treated cells were also used as negative controls for flow cytometry. Cells were washed twice before collection and subjected to processing using the Click-iT Flow Cytometry Assay kit (#C10418, Invitrogen) following the manufacturer’s instructions. Labeled cells were analyzed using a BD LSR Fortessa instrument and graphed as Alexa Fluor 647 (AF647) Mean Fluorescence Intensity (normalized to DMSO control treated with OP-Puro).
Minuesa G., Albanese S.K., Xie W., Kazansky Y., Worroll D., Chow A., Schurer A., Park S.M., Rotsides C.Z., Taggart J., Rizzi A., Naden L.N., Chou T., Gourkanti S., Cappel D., Passarelli M.C., Fairchild L., Adura C., Glickman J.F., Schulman J., Famulare C., Patel M., Eibl J.K., Ross G.M., Bhattacharya S., Tan D.S., Leslie C.S., Beuming T., Patel D.J., Goldgur Y., Chodera J.D, & Kharas M.G. (2019). Small-molecule targeting of MUSASHI RNA-binding activity in acute myeloid leukemia. Nature Communications, 10, 2691.
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2

Measuring Protein Synthesis in HSCs

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Protein synthesis rate was assessed by O-Propargyl-puromycin (OP-Puro) as described previously(Vu et al., 2017b (link)). Mettl3 f/f and Mettl3 cKO HSC cells were sorted and treated with 50 μM O-propargyl-puromycin (OP-Puro; NU-931–05, Jena Bioscience) for 1 hour. Cells were washed once, collected, and then subjected to processing using the Click-iT Flow Cytometry Assay kit (C10418, Invitrogen) following the manufacturer’s instructions. Labeled cells were analyzed using a BD Fortessa instrument.
Cheng Y., Luo H., Izzo F., Pickering B.F., Nguyen D., Myers R., Schurer A., Gourkanti S., Brϋning J.C., Vu L.P., Jaffrey S.R., Landau D.A, & Kharas M.G. (2019). m6A RNA Methylation Maintains Hematopoietic Stem Cell Identity and Symmetric Commitment. Cell reports, 28(7), 1703-1716.e6.
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3

Protein Synthesis Measurement by Flow Cytometry

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Cells were plated at 100,000 cells/ml density and treated with 50 uM OP-Puro (NU-931-05, Jena Bioscience). Control cells were treated with cyclohexamide (CHX) at 150 ug/ml for 15 minute. Cells were washed twice prior to collection and subjected to processed using Click-iT® Flow Cytometry Assay Kit (C10418 – Invitrogene) following the manufacturer’s instructions. Labelled cells were analyzed using BD Fortessa instrument.
Vu L.P., Prieto C., Amin E.M., Chhangawala S., Krivtsov A., Calvo-Vidal M.N., Chou T., Chow A., Minuesa G., Park S.M., Barlowe T.S., Taggart J., Tivnan P., Deering R.P., Chu L.P., Kwon J.A., Meydan C., Perales-Paton J., Arshi A., Gönen M., Famulare C., Patel M., Paietta E., Tallman M.S., Lu Y., Glass J., Garret-Bakelman F., Melnick A., Levine R., Al-Shahrour F., Järås M., Hacohen N., Hwang A., Garippa R., Lengner C.J., Armstrong S.A., Cerchietti L., Cowley G.S., Root D., Doench J., Leslie C., Ebert B.L, & Kharas M.G. (2017). Functional screen of MSI2 interactors identifies an essential role for SYNCRIP in myeloid leukemia stem cells. Nature genetics, 49(6), 866-875.
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4

Protein Synthesis Measurement by Flow Cytometry

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Cells were plated at 100,000 cells/ml density and treated with 50 uM OP-Puro (NU-931-05, Jena Bioscience). Control cells were treated with cyclohexamide (CHX) at 150 ug/ml for 15 minute. Cells were washed twice prior to collection and subjected to processed using Click-iT® Flow Cytometry Assay Kit (C10418 – Invitrogene) following the manufacturer’s instructions. Labelled cells were analyzed using BD Fortessa instrument.
Vu L.P., Prieto C., Amin E.M., Chhangawala S., Krivtsov A., Calvo-Vidal M.N., Chou T., Chow A., Minuesa G., Park S.M., Barlowe T.S., Taggart J., Tivnan P., Deering R.P., Chu L.P., Kwon J.A., Meydan C., Perales-Paton J., Arshi A., Gönen M., Famulare C., Patel M., Paietta E., Tallman M.S., Lu Y., Glass J., Garret-Bakelman F., Melnick A., Levine R., Al-Shahrour F., Järås M., Hacohen N., Hwang A., Garippa R., Lengner C.J., Armstrong S.A., Cerchietti L., Cowley G.S., Root D., Doench J., Leslie C., Ebert B.L, & Kharas M.G. (2017). Functional screen of MSI2 interactors identifies an essential role for SYNCRIP in myeloid leukemia stem cells. Nature genetics, 49(6), 866-875.
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5

Measuring Protein Synthesis in HSCs

Cited 1 time
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Protein synthesis rate was assessed by O-Propargyl-puromycin (OP-Puro) as described previously(Vu et al., 2017b (link)). Mettl3 f/f and Mettl3 cKO HSC cells were sorted and treated with 50 μM O-propargyl-puromycin (OP-Puro; NU-931–05, Jena Bioscience) for 1 hour. Cells were washed once, collected, and then subjected to processing using the Click-iT Flow Cytometry Assay kit (C10418, Invitrogen) following the manufacturer’s instructions. Labeled cells were analyzed using a BD Fortessa instrument.
Cheng Y., Luo H., Izzo F., Pickering B.F., Nguyen D., Myers R., Schurer A., Gourkanti S., Brϋning J.C., Vu L.P., Jaffrey S.R., Landau D.A, & Kharas M.G. (2019). m6A RNA Methylation Maintains Hematopoietic Stem Cell Identity and Symmetric Commitment. Cell reports, 28(7), 1703-1716.e6.
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6

Cell Proliferation Assay with EdU

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To assess in vitro proliferation, cells were analyzed with the Click-iT Flow Cytometry Assay Kit (Invitrogen; C10419) according to the manufacturer’s protocol, using a labeling time of 4 hours at an EdU concentration of 10 µg/ml.
Hutter K., Rülicke T., Szabo T.G., Andersen L., Villunger A, & Herzog S. (2022). The miR-15a/16-1 and miR-15b/16-2 clusters regulate early B cell development by limiting IL-7 receptor expression. Frontiers in Immunology, 13, 967914.
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7

EdU Labeling of Neurospheres

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For 5-ethynyl-2′-deoxyuridine (EdU) (Invitrogen) labeling of neurospheres in culture, cells were incubated in neurospheres media containing 10 µM EdU for 12 h and then processed for EdU detection according to the protocol of the manufacturer (Click-IT Flow Cytometry Assay kit; Invitrogen).
Pan L., North H.A., Sahni V., Jeong S.J., Mcguire T.L., Berns E.J., Stupp S.I, & Kessler J.A. (2014). β1-Integrin and Integrin Linked Kinase Regulate Astrocytic Differentiation of Neural Stem Cells. PLoS ONE, 9(8), e104335.
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