Nexin reagent

Manufactured by Merck Group

Sourced in United States

The Nexin reagent is a laboratory product manufactured by the Merck Group. It is a chemical reagent used in various analytical and research applications. The core function of the Nexin reagent is to facilitate specific chemical reactions and analyses, but a detailed description of its intended use cannot be provided in an unbiased and factual manner without the risk of extrapolation.

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Guava Nexin Reagent (110 citations) Guava Nexin Reagent kit (4 citations) Guava Cell Nexin Reagent (2 citations) Guava Nexin Reagent for cell apoptosis kit (2 citations)

12 protocols using nexin reagent

Annexin V and PI Staining for Jurkat Cell Viability

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Jurkat cells were washed, counted and diluted to a density of 1 × 10⁵cells/ml in PBS. For each sample, 100 uL of cells in suspension was mixed with 100 ul of room-temperature annexin V-PE staining reagent (Guava Nexin Reagent) and incubated for 20 minutes at room temperature in the dark. After incubation, samples were acquired using a Guava EasyCyte Mini flow cytometer.
Cell viability was assessed using propidium iodide staining. To 200 ul of live cells in suspension PI solution was added to final concentration 10 ug/ml. Samples were incubated for 5 minutes at room temperature in the dark. After incubation, samples were acquired using a Guava EasyCyte Mini flow cytometer.

Kaminski R., Chen Y., Fischer T., Tedaldi E., Napoli A., Zhang Y., Karn J., Hu W, & Khalili K. (2016). Elimination of HIV-1 Genomes from Human T-lymphoid Cells by CRISPR/Cas9 Gene Editing. Scientific Reports, 6, 22555.

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Apoptosis Measurement by Flow Cytometry

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Apoptotic fraction was evaluated by flow cytometry using the Guava Nexin assay (Guava Technologies, Hayward, CA). Cells were exposed LB100 (2.5 μM) for 4 hours prior to administration of 5 Gy or sham radiation. Cells were trypsinized and stained per manufacturer's instructions with Nexin Reagent to assess annexin-V conjugated to phycoerithrin as a marker of cells in early apoptosis and 7-AAD as an indicator of late apoptosis (Guava Technologies). Analysis was performed on an EasyCyte Plus flow cytometer (Guava Technologies).

Gordon I.K., Lu J., Graves C.A., Huntoon K., Frerich J.M., Hanson R.H., Wang X., Hong C.S., Ho W., Feldman M.J., Ikejiri B., Bisht K., Chen X.S., Tandle A., Yang C., Arscott W.T., Ye D., Heiss J.D., Lonser R.R., Camphausen K, & Zhuang Z. (2015). Protein phosphatase 2A inhibition with LB100 enhances radiation-induced mitotic catastrophe and tumor growth delay in glioblastoma. Molecular cancer therapeutics, 14(7), 1540-1547.

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Apoptosis Evaluation by Flow Cytometry

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Apoptotic fraction was evaluated by flow cytometry using the Guava Nexin assay (Guava Technologies, Hayward, CA). Cells were exposed LB100 (2.5 μM) for 3 hours prior to administration of 8 Gy or sham radiation. Cells were trypsinized and stained per manufacturer's instructions with Nexin Reagent to assess annexin-V conjugated to phycoerythrin as a marker of cells in early apoptosis and 7-AAD as an indicator of late apoptosis (Guava Technologies). Analysis was performed on an EasyCyte Plus flow cytometer.

Lv P., Wang Y., Ma J., Wang Z., Li J.L., Hong C.S., Zhuang Z, & Zeng Y.X. (2014). Inhibition of protein phosphatase 2A with a small molecule LB100 radiosensitizes nasopharyngeal carcinoma xenografts by inducing mitotic catastrophe and blocking DNA damage repair. Oncotarget, 5(17), 7512-7524.

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Apoptosis Evaluation of BMMSCs

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BMMSCs derived from ZL and ZDF rats (passage 2) were cultured in serum-free αMEM for 48 h. The percentage of apoptotic cells was then determined using Guava^® Nexin Reagent and flow cytometry analysis according to the manufacturer’s instruction.

Ribot J., Denoeud C., Frescaline G., Landon R., Petite H., Pavon-Djavid G., Bensidhoum M, & Anagnostou F. (2021). Experimental Type 2 Diabetes Differently Impacts on the Select Functions of Bone Marrow-Derived Multipotent Stromal Cells. Cells, 10(2), 268.

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Nexin Reagent Flow Cytometry

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Nexin reagent (Millipore #4500-0455) by manufacturer’s instructions using the Guava EasyCyte flow cytometer.

Amin A.D., Rajan S.S., Liang W.S., Pongtornpipat P., Groysman M.J., Tapia E.O., Peters T.L., Cuyugan L., Adkins J., Rimsza L.M., Lussier Y.A., Puvvada S.D, & Schatz J.H. (2015). Evidence Suggesting that Discontinuous Dosing of ALK Kinase Inhibitors May Prolong Control of ALK+ Tumors. Cancer research, 75(14), 2916-2927.

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Multiparametric Flow Cytometry of Apoptosis

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Apoptosis was measured with the Guava EasyCytePlus using Nexin reagent per the manufacturer's protocol (Millipore). Cell cycle, DNA damage and apoptosis were measured simultaneously using the Apoptosis, DNA damage and Cell Proliferation Kit (BD Biosciences) according to the manufacturer's instructions. Fixation and permeabilization buffers from this kit were used to prepare cells for staining with PE-linked human CD45 and phopho-CDK1 antibodies followed by secondary staining with anti-rabbit Alexa Fluor 488. Stained cells were analyzed on a Gallios 561 flow cytometer (Beckman Coulter).

Ford J.B., Baturin D., Burleson T.M., Van Linden A.A., Kim Y.M, & Porter C.C. (2015). AZD1775 sensitizes T cell acute lymphoblastic leukemia cells to cytarabine by promoting apoptosis over DNA repair. Oncotarget, 6(29), 28001-28010.

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Quantifying Apoptosis by Flow Cytometry

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Annexin V/7-AAD staining was conducted on living cells using the Nexin reagent (Millipore). The percentage of apoptotic cells was determined by flow cytometry using the Guava system (Millipore). For quantification of Annexin V-positive, or Annexin V/7-AAD-positive cells, the same gate settings were applied for all samples. For each experiment, at least three biological replica were analyzed. Asterisks represent the significance that was calculated using Student's t-test.

Kramer D., Schön M., Bayerlová M., Bleckmann A., Schön M.P., Zörnig M, & Dobbelstein M. (2015). A pro-apoptotic function of iASPP by stabilizing p300 and CBP through inhibition of BRMS1 E3 ubiquitin ligase activity. Cell Death & Disease, 6(2), e1634-.

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Flow Cytometry-based Cell Viability Assessment

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Cell viability and proliferation assays were evaluated on a Guava easyCyte HT System Flow Cytometer (Millipore, Billerica, MA, USA) using ViaCount reagent (4000-0040) for analysis of viability and cell number and NEXIN reagent (4000-0450) for analysis of early and late apoptotic cells according to the manufacturer's instructions (Millipore). Parameters were set using untreated cells and analyzed in triplicate.

Margaryan N.V., Gilgur A., Seftor E.A., Purnell C., Arva N.C., Gosain A.K., Hendrix M.J, & Strizzi L. (2016). Melanocytes Affect Nodal Expression and Signaling in Melanoma Cells: A Lesson from Pediatric Large Congenital Melanocytic Nevi. International Journal of Molecular Sciences, 17(3), 418.

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Cell Viability and Apoptosis Assay

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Cells were seeded at 0.5–1.0 × 10⁵/mL in triplicate wells of 48-well tissue culture plates. Where indicated, the cells were treated with drug for a period of 48–72 hours. After treatment, a sample of cells from each well was stained with propidium iodide (PI; 10 µg/mL) and viable cells (PI⁻) were counted with a flow cytometer (Guava easyCyte 8HT). Alternatively, cells were stained using 7-aminoactinomycin D/anti-Annexin V (Nexin reagent, Millipore EMD) to detect apoptotic cells.

Gregory M.A., Nemkov T., Reisz J.A., Zaberezhnyy V., Hansen K.C., D’Alessandro A, & DeGregori J. (2017). Glutaminase inhibition improves FLT3 inhibitor therapy for acute myeloid leukemia. Experimental hematology, 58, 52-58.

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Annexin V Apoptosis Assay

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Cells (1 × 10⁶/mL) were treated with various concentrations of LtxA for indicated times. Following treatment, 100 µL of treated cells were transferred to 96-well black bottom plates. An aliquot of 100 µL of Nexin reagent (Millipore) was then added to each well containing 100 µL of cells. The plate was incubated in the dark and on a shaker for 10 min. Following incubation, the luminescence was read using a microtiter plate reader (BioTek Synergy 2). Treatments were performed in triplicate and 5000–10,000 events were recorded. Apoptosis was determined as the percentage of Annexin V⁺ cells relative to an untreated cell control.

Prince D.J., Patel D, & Kachlany S.C. (2021). Leukotoxin (LtxA/Leukothera) induces ATP expulsion via pannexin-1 channels and subsequent cell death in malignant lymphocytes. Scientific Reports, 11, 18086.

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