Flow cytometer facscalibur dxp

Manufactured by BD

Sourced in China, United States

The BD FACSCalibur DXP is a flow cytometer instrument designed for cell analysis and sorting. It utilizes laser-based technology to detect and measure various physical and fluorescent characteristics of cells or particles suspended in a fluid stream.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (7 mentions) Cck 8 reagent, by Beyotime (2 mentions) Clickredu solution, by Thermo Fisher Scientific (2 mentions) Infinite m200, by Tecan (1 mentions) 5 ethynyl 2 deoxyuridine edu assay kit, by RiboBio (1 mentions) Anti phospho histone h2a x ser139 antibody, by Merck Group (1 mentions) Alexafluor 488 labelled goat anti mouse secondary antibody, by Thermo Fisher Scientific (1 mentions) Mtt assay, by Roche (1 mentions)

Spelling variants of this product

FACSCalibur (18538 citations) FACSCalibur flow cytometer (10214 citations) FACSCalibur cytometer (1030 citations) FACSCalibur flow (29 citations)

7 protocols using flow cytometer facscalibur dxp

Quantifying Cell Proliferation via CCK-8 and EdU Assays

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Approximately 1.0 × 10⁴ transfected SCC-4 and CAL-27 cells were cultured in 96-well plates, and then underwent 1-h incubation with CCK-8 reagent (Beyotime, Shanghai, China). The absorbance at 450 nm was recorded using an Infinite M200 multimode microplate reader (Tecan, Shanghai, China).
After approximately 48-h transfection, the 5-ethynyl-2’-deoxyuridine (EdU) assay kit provided by Ribo (Guangzhou, China) was utilized to examine the proliferation of SCC-4 and CAL-27 cells. Specifically, cells were grown in culture medium containing EdU (Invitrogen) solution (1:1,000). At the proliferative stage, the cells were labeled with EdU for 2 h, followed by rinsing with PBS (0.5 g/ml) thrice. Subsequently, the cells were stained by 4′,6-diamidino-2-phenylindole (DAPI) from Invitrogen for 10 min at indoor temperature in the dark and underwent PBS rinsing more than twice. Ultimately, assessment of the stained cells was implemented via the FACSCalibur DxP flow cytometer (BD Biosciences, Shanghai, China).

Ai Y., Wei H., Wu S., Tang Z., Li X, & Zou C. (2021). Exosomal LncRNA LBX1-AS1 Derived From RBPJ Overexpressed-Macrophages Inhibits Oral Squamous Cell Carcinoma Progress via miR-182-5p/FOXO3. Frontiers in Oncology, 11, 605884.

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Cell Proliferation Assays in Glioblastoma

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Approximately, 1.0×10³ transfected U87 MG and U251 cells were cultured in 96-well plates. Cell Counting Kit-8 (CCK-8; 10 µl) reagent (Beyotime Institute of Biotechnology) was added and incubated at 37°C for 1 h. The absorbance at 450 nm was recorded using an Infinite M200 multimode microplate reader (Tecan Group, Ltd.).
After approximately 48 h of transfection, the 5-ethynyl-2´-deoxyuridine (EdU) assay kit provided by Guangzhou Ribo Co., Ltd., was used to examine the proliferation of U87 MG and U251 cells. Specifically, cells were grown in culture medium containing EdU (cat. no. A10044; Invitrogen; Thermo Fisher Scientific, Inc.) solution (1,000:1). At the proliferative stage, the cells were labeled with EdU for 2 h, followed by three rinses with phosphate-buffered saline (PBS; 0.5 g/ml). Subsequently, 4′,6-diamidino-2-phenylindole (DAPI; Invitrogen; Thermo Fisher Scientific, Inc.) was used to stain nuclei of the washed cells for 10 min at room temperature in the dark. The DAPI-stained cells were washed more than twice with PBS. Stained cells were analyzed using the FACSCalibur DxP flow cytometer (BD Biosciences).

Dai Y., Zhang Y., Hao M, & Zhu R. (2021). LINC00665 functions as a competitive endogenous RNA to regulate AGTR1 expression by sponging miR-34a-5p in glioma. Oncology Reports, 45(3), 1202-1212.

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Evaluating Proliferation in Glioblastoma Cells

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Approximately 1.0 × 10³ transfected U87 MG and U373 MG cells were cultured in 96-well plates, and then underwent 1 h incubation with CCK-8 reagent (Beyotime, Shanghai, China). The absorbance at 450 nm was recorded using an Infinite M200 multimode microplate reader (Tecan, Shanghai, China).
After approximately 48 h transfection, the 5-ethynyl-2’-deoxyuridine (EdU) assay kit provided by Ribo (Guangzhou, China) was utilized to examine the proliferation of U87 MG and U373 MG cells. Specifically, cells were grown in culture medium containing EdU (Invitrogen) solution (1:1000). At the proliferative stage, the cells were labeled with EdU for 2 h, followed by rinsing with PBS (0.5 g/mL) thrice. Subsequently, the cells were stained by 4′,6-diamidino-2-phenylindole (DAPI) from Invitrogen for 10 min at indoor temperature in the dark and underwent PBS rinsing more than twice. Ultimately, assessment of the stained cells was implemented via the FACSCalibur DxP flow cytometer (BD Biosciences, Shanghai, China).

Shi L., Cao Y., Yuan W., Guo J, & Sun G. (2022). Exosomal circRNA BTG2 derived from RBP-J overexpressed-macrophages inhibits glioma progression via miR-25-3p/PTEN. Cell Death & Disease, 13(5), 506.

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EdU Proliferation Assay for U87 and U251 Cells

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After transfection of pcDNA/HCG11 and pcDNA‐NC for 48 hours, the proliferation abilities of U87 and U251 cells were detected by EdU cell proliferation (Ribo, Guangzhou, China). Briefly, the ClickREdU solution (Invitrogen) was added to the culture medium (1000:1). The cells which were in proliferating phase were tabbed with EdU for 2 hours. Next, the cells were washed for three times with 0.5 g/mL of PBS. Subsequently, DAPI (Invitrogen) nuclei counterstained the washed cells for 10 minutes at normal temperature in the dark place. The DAPI‐marked cells were then washed more than twice with PBS. Finally, all marked cells were analysed and measured by the flow cytometer FACSCalibur DxP (BD Biosciences, Franklin Lakes, NJ, USA).

Chen Y., Bao C., Zhang X., Lin X., Huang H, & Wang Z. (2019). Long non‐coding RNA HCG11 modulates glioma progression through cooperating with miR‐496/CPEB3 axis. Cell Proliferation, 52(5), e12615.

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DNA Damage Assessment in DU145 Cells

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Time course DU145-Parental control and DU145-Parental-UCA1 siRNA samples were fixed in 70% ethanol on ice for 30 min. After washing with PBS, cell pellets were blocked in 0.1%(v/v) Triton X-100 and 1% (v/v) goat serum at room temperature for 1 hour, and then incubated with anti-phosphohistone H2AX (ser-139) antibody (EMD Millipore, MA, USA) overnight at 4°C. Following three washes with PBS, cells were incubated with Alexafluor 488 labelled goat-anti-mouse secondary antibody (Invitrogen, Ontario, Canada) at room temperature in the dark for 1 hour. Cells were washed three times with PBS and nuclei counterstained with 0.5 g/mL DAPI (Invitrogen, Ontario, Canada) in PBS for 10 min in the dark at room temperature, followed by three washes with PBS. Samples were analyzed using flow cytometer FACSCalibur DXP (BD Biosciences).

Ghiam A.F., Taeb S., Huang X., Huang V., Ray J., Scarcello S., Hoey C., Jahangiri S., Fokas E., Loblaw A., Bristow R.G., Vesprini D., Boutros P, & Liu S.K. (2016). Long non-coding RNA urothelial carcinoma associated 1 (UCA1) mediates radiation response in prostate cancer. Oncotarget, 8(3), 4668-4689.

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Evaluating miR-590-3p Effects on Colorectal Cancer Cell Proliferation

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After transfecting miR-590-3p mimics and negative control for 48h, 3,000 cells per well were grew in 96-well plates for 6 h (n=5). Then the proliferation of HCT116 cells or SW480 cells were detected by MTT assay (Roche, Basel, Switzerland) and EdU cell proliferation (Ribo, Guangzhou, China).
The transfected cells were continuously cultured for 1d, 2d, 3d, 4d and 5d, further added 10 μL MTT for 4 h, 37°C. Then removed the medium, the well were added DMSO for 10 min. The OD values were determined at 490nm by a microplate reader. In addition, the ClickREdU solution (Invitrogen, Carlsbad, CA, USA) was added to the culture medium at a ratio of 1000: 1 and the cells, which were in proliferating phase, were marked with EdU for 2 h. Similarly, the transfected cells were washed three times with PBS 0.5 g/mL DAPI (Invitrogen, Ontario, Canada) nuclei counterstained the washed cells for 10 min at room temperature in a dark. Then the DAPI-marked cells were washed three times with PBS. All marked cells were analyzed by flow cytometer FACSCalibur DXP (BD Biosciences).

Sun Z.Q., Shi K., Zhou Q.B., Zeng X.Y., Liu J., Yang S.X., Wang Q.S., Li Z., Wang G.X., Song J.M., Yuan W.T, & Wang H.J. (2017). MiR-590-3p promotes proliferation and metastasis of colorectal cancer via Hippo pathway. Oncotarget, 8(35), 58061-58071.

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Cellular Proliferation Assay Using EdU

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The proliferation ability of transfected cells was evaluated by applying EdU cell proliferation (Ribo, Guangzhou, China). Briefly, cells in the proliferating phase were treated with EdU for 2 h. After the cells were washed three times with 0.5 g/ml of PBS, DAPI (Invitrogen) nuclei counterstained cells for 10 min at room temperature in a dark room. Cells marked by DAPI were washed three times with PBS. At length, the number of marked cells were measured with the flow cytometer FACSCalibur DXP (BD Biosciences, Franklin Lakes, NJ, USA).

Wu D.M., Wang S., Wen X., Han X.R., Wang Y.J., Shen M., Fan S.H., Zhang Z.F., Shan Q., Li M.Q., Hu B., Lu J., Chen G.Q, & Zheng Y.L. (2018). LncRNA SNHG15 acts as a ceRNA to regulate YAP1-Hippo signaling pathway by sponging miR-200a-3p in papillary thyroid carcinoma. Cell Death & Disease, 9(10), 947.

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