Dxflex

Manufactured by Beckman Coulter

Sourced in United States, China

The DxFLEX is a flow cytometry instrument developed by Beckman Coulter. It is designed to analyze and sort cells based on their physical and fluorescent characteristics. The DxFLEX utilizes advanced optics and software to provide high-performance cell analysis and sorting capabilities.

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Lab products found in correlation

Cytexpert for dxflex, by Beckman Coulter (3 mentions) Cytexpert software, by Beckman Coulter (3 mentions) Annexin 5 fitc, by BD (2 mentions) Prism 9, by GraphPad (2 mentions) Anti cd11c pe, by BioLegend (1 mentions) Anti cd80 pe, by BioLegend (1 mentions) Anti cd11c fitc, by BioLegend (1 mentions) Annexin 5 fluorescein isothiocyanate fitc kit, by BD (1 mentions) Ab19898, by Abcam (1 mentions) Ab205718, by Abcam (1 mentions) Trypan blue stain, by Thermo Fisher Scientific (1 mentions) Percoll, by Cytiva (1 mentions) Anti mouse cd16 32, by Thermo Fisher Scientific (1 mentions) Anti mouse cd3 fitc, by BD (1 mentions) Anti mouse nk1.1 apc, by BD (1 mentions) Fitc conjugated monoclonal antibodies, by BioLegend (1 mentions) Cd105, by BioLegend (1 mentions) Pe annexin 5 apoptosis detection kit, by BD (1 mentions) Annexin 5 apc pi apoptosis kit, by MultiSciences Biotech (1 mentions) Cd44 pe, by BD (1 mentions)

Spelling variants of this product

DxFLEX flow cytometer (85 citations) Beckman DxFLEX (5 citations) DxFLEX flow cytometry system (5 citations) CytExpert for DxFLEX software (4 citations) DxFlex ow cytometer (3 citations) DxFLEX ow cytometry system (2 citations) CytExpert for DxFLEX version 1.0 (2 citations) DxFlex system (2 citations) DxFLEX ow cytometry (2 citations) Beckman DxFLEX FCM (1 citations)

87 protocols using dxflex

Quantifying Reactive Oxygen Species in Cells

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Cells were stained with the following monoclonal antibodies: anti-CD45.2 (clone 104), anti-CD11b (clone M1/70), anti-Ly6G (clone 1A8), and anti-MPO (clone EPR20257). We acquired cells on a Beckman DxFlex and analyzed with CytExpert for DxFlex.
The content of ROS in cells was detected using a DHE fluorescent probe. DHE is one of the most common fluorescent detection probes for superoxide anions. Cells were incubated in the dark with DHE solution for 30 min at 37 °C. Cells were washed with PBS and then stained with the before mentioned monoclonal antibodies for 30 min at 4 °C. Cells were washed with PBS and resuspended with PBS. Finally, we acquired cells on a Beckman DxFlex and analyzed with CytExpert for DxFlex. Detailed methods are outlined in one of our previous studies [27 (link)].

Han F., Ding Z.F., Shi X.L., Zhu Q.T., Shen Q.H., Xu X.M., Zhang J.X., Gong W.J., Xiao W.M., Wang D., Chen W.W., Hu L.H, & Lu G.T. (2023). Irisin inhibits neutrophil extracellular traps formation and protects against acute pancreatitis in mice. Redox Biology, 64, 102787.

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Antigen Uptake and Presentation in BMDCs

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FITC-labeled OVA and FITC-OVA@SVMAV were used for antigen uptake experiment. BMDCs were plated in 24-well plates at 2× 10⁵ per well with BMDC complete medium, and then treated with different combinations of drugs for 4 hours at the dose of 40 nmol/mL for each compound and maintained without any treatments for 20 hours. BMDCs were then collected and stained with anti-CD11c-PE (#117307, BioLegend, USA). The percentage of FITC-positive CD11c⁺ DCs was analyzed by a flow cytometer (DxFLEX, Beckman Coulter, USA).
Unlabeled OVA and OVA@SVMAV were used for antigen presentation experiment. BMDCs (2×10⁵ cells/well) were incubated with different combinations of drugs for 4 hours at the dose of 40 nmol/mL for each compound and maintained without any treatments for 20 hours. Cells were collected and stained with anti-CD11c-FITC (#117305, BioLegend, USA), anti-CD80-PE (#104707, BioLegend, USA), and anti-SIINFEKL/H-2Kb-APC (#141606, BioLegend, USA) for 30 min at room temperature and subject for flow cytometry analysis (DxFLEX, Beckman Coulter, USA).

Zhang L., Huang J., Chen X., Pan C., He Y., Su R., Guo D., Yin S., Wang S., Zhou L., Chen J., Zheng S, & Qiao Y. (2021). Self-assembly nanovaccine containing TLR7/8 agonist and STAT3 inhibitor enhances tumor immunotherapy by augmenting tumor-specific immune response. Journal for Immunotherapy of Cancer, 9(8), e003132.

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Flow Cytometric Characterization of CD133-Expressing Cells

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After cell treatment and counting, the cells were resuspended with PBS containing 2% BSA to reach a cell concentration of 1 × 10⁶ cells/ml. No antibody was added to the blank control tube, with 5–20 μl rabbit antibody to CD133 (ab19898, Abcam) subsequently added to the sample tubes, followed by mixing and incubation at room temperature for 30 min. The cells were centrifuged at 1000 rpm for 5 min, and subsequently resuspended with 100 μl PBS. Fluorescence-labeled secondary antibody, namely, goat anti-rabbit to immunoglobulin G (IgG) H&L (HRP) (ab205718, Abcam) was added to the cells and incubated at room temperature under dark conditions for 30 min. Following centrifugation at 1000 rpm for 5 min, the cells were resuspended with 100–200 μl PBS, with different samples examined using a flow cytometer (DxFLEX, Beckman Coulter Inc., Chaska, MN, USA).
As per the manual, cell apoptosis was detected by Annexin V-Fluorescein Isothiocyanate (FITC) kit (BD Bioscience, NY, USA). Briefly, cells were detached by trypsin, collected, washed with cold PBS and centrifuged following the culmination of a 3-day transfection. The cell precipitate was resuspended in cold binding buffer solution and added with Annexin V-FITC (1.25 μl/0.5 ml) and propidium iodide (10 μl/0.5 ml). Flow cytometer (DxFLEX, Beckman) was employed for detection purposes. Three duplicates were set for each well.

Peng L., Fu J., Chen Y., Ming Y., He H., Zeng S., Zhong C, & Chen L. (2022). Transcription factor SNAI2 exerts pro-tumorigenic effects on glioma stem cells via PHLPP2-mediated Akt pathway. Cell Death & Disease, 13(6), 516.

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Cellular Fluorescence Analysis of Saos-2 Cells

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The Saos-2 cells exposed to LDOX or free DOX based on the IC₅₀ values and incubated under standard conditions for 48 h were analyzed for their cellular fluorescence in a DxFlex flow cytometer (Beckman Coulter, Brea, CA, USA) using the CytExpert v.2.5 software. The fluorescence intensity was measured from the ECD channel [91 (link)]. The gating strategy used FSC-A and SSC-A to detect viable, single-cell events. A total of at least 25,000 events were collected from each sample.

Orel V.E., Diedkov A.G., Ostafiichuk V.V., Lykhova O.O., Kolesnyk D.L., Orel V.B., Dasyukevich O.Y., Rykhalskyi O.Y., Diedkov S.A, & Prosvietova A.B. (2024). Combination Treatment with Liposomal Doxorubicin and Inductive Moderate Hyperthermia for Sarcoma Saos-2 Cells. Pharmaceuticals, 17(1), 133.

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Murine Liver Lymphocyte Isolation

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After the mouse liver was removed, a mononuclear cell suspension was prepared by mechanical grinding. Percoll (Cytiva, 17,089,101, Sweden) was used for lymphocyte gradient separation. The cells were washed twice in PBS buffer containing 0.2% bovine serum albumin (BSA) and resuspended in PBS buffer for counting cell number with Trypan blue stain (Gibco, 15250-061, USA). Anti-mouse CD16/32 (eBioscience, 14-0161-86, United States) was used to block the Fc receptor. The following antibodies were used for immunophenotyping analysis of the samples: anti-mouse CD3-FITC (BD, 555,274, USA), anti-mouse NK1.1-APC (BD, 550,627, USA), and anti-mouse NKG2D-PE-Cy7(BD, 562,614, USA), Data were obtained by the flow cytometry (DxFLEX, Beckman Coulter, USA) and analysed using FlowJo 10.0 software.

Ping D.B., Sun X., Peng Y, & Liu C.H. (2023). Cyp4a12-mediated retinol metabolism in stellate cells is the antihepatic fibrosis mechanism of the Chinese medicine Fuzheng Huayu recipe. Chinese Medicine, 18, 51.

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Immunophenotypic Characterization of ADSCs

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Approximately 1 × 10⁶ ADSCs from the third passage were fixed in 4% paraformaldehyde (PFA) for 15 min. ADSCs and TADSCs were incubated with phycoerythrin (PE)- or fluorescein isothiocyanate (FITC)-conjugated monoclonal antibodies against CD29 (BioLegend, San Diego, CA, United States), CD44 (BioLegend, San Diego, CA, United States), CD45 (BioLegend, San Diego, CA, United States), and CD105 (BioLegend, San Diego, CA, United States) at room temperature (RT) in the dark for 1 h. Subsequently, both ADSCs and TADSCs were examined by flow cytometry (DxFLEX, Beckman Coulter, Brea, CA, United States). Monoclonal antibodies, including CD29-PE, CD44-PE, and CD105-PE, were used to identify the mesenchymal lineage. Simultaneously, hematopoietic and angiogenic phenotypes were excluded using CD45-FITC antibodies.

Zhang G., Song S., Chen Z., Liu X., Zheng J., Wang Y., Chen X, & Song Y. (2024). Inhibition of PTEN promotes osteointegration of titanium implants in type 2 diabetes by enhancing anti-inflammation and osteogenic capacity of adipose-derived stem cells. Frontiers in Bioengineering and Biotechnology, 12, 1358802.

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Sepsis Biomarkers and Immune Profiles

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T lymphocyte subsets were measured using a DxFLEX flow cytometer from Beckman, the United States of America. The model of ventilator used was Philips V60. The results of sepsis biomarkers and immune cells were obtained within 24 h of patient admission.

Zhang Y., Wang J., Hu L., Xuan J., Qu Y., Li Y., Ye X., Yang L., Yang J., Zhang X., Wang J, & Wei B. (2021). Predictive Value of Immune Cell Subsets for Mortality Risk in Patients With Sepsis. Clinical and Applied Thrombosis/Hemostasis, 27, 10760296211059498.

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TGF-β1-Induced Apoptosis Assay

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JS1 cells were incubated with TGF-β1 (10 ng/ml) for 12 h and pre-treated with or without 10 µM SB203580 (cat. no. ab120162; Abcam) for 1 h. In the absence or presence of the previously mentioned agents, JS1 cells were labeled with PE Annexin V and 7-amino-actino-mycin using a PE Annexin V Apoptosis Detection kit according to the manufacturer's protocol (BD Biosciences) and analyzed using flow cytometry (DxFLEX; Beckman Coulter, Inc.). Flow cytometry data were analyzed using CytExpert software (version 2.0; Beckman Coulter, Inc.).

Zhang J., Jiang N., Ping J, & Xu L. (2020). TGF-β1-induced autophagy activates hepatic stellate cells via the ERK and JNK signaling pathways. International Journal of Molecular Medicine, 47(1), 256-266.

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Circadian Regulator circ-ATXN2 Modulates Apoptosis

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To observe the effects of circ-ATXN2 overexpression on cell death, CoCl₂ was used to create apoptotic models of the cells. Vector-GFP- and circ-ATXN2–GFP-expressing cells were seeded at 1 × 10⁵ cells per well in 6-well plates and incubated with 500 μM CoCl₂ for 24 h. Annexin V-APC/PI apoptosis kit (MULTI SCIENCES) was used to determine cell apoptosis. Cells were collected in 500 μl of binding buffer with 3 μl Annexin V-APC and 3 μl PI for 20 min at room temperature in the dark. Apoptotic cells were detected using a flow cytometer (DxFLEX, Beckman Coulter, United States) (He et al., 2021 (link); Song et al., 2021 (link)).

Song X.H., He N., Xing Y.T., Jin X.Q., Li Y.W., Liu S.S., Gao Z.Y., Guo C., Wang J.J., Huang Y.Y., Hu H, & Wang L.L. (2021). A Novel Age-Related Circular RNA Circ-ATXN2 Inhibits Proliferation, Promotes Cell Death and Adipogenesis in Rat Adipose Tissue-Derived Stromal Cells. Frontiers in Genetics, 12, 761926.

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Immune Cell Phenotyping Protocol

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Hemolysin, IL‐23, and flow cytometry antibodies of CD4‐FITC, CD44‐PE, and CD62L‐PE‐CF59 were purchased from Becton, Dickinson and Company, USA. We used the DXflex flow cytometer (Beckman Coulter).

Zhou J., Du X., Abulajiang X., Geli W., Pu X., Tailaiti S., Lin J., Li Y, & Ye J. (2023). The role of memory T cells in Echinococcus granulosus‐induced sensitization. Immunity, Inflammation and Disease, 11(8), e948.

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