LN229 and U87MG cells with logarithmic growth phase were plated into a 6-well plate and the cells were treated with different concentrations of S4. Then PBS was used to wash the collected cells. Glioma cells were stained with annexin V-FITC and propidium iodide (PI) according to the to the manufacturer’s instructions (Keygen Biotech/KGA108), and the cells were detected on the flow cytometer within 1 h. Three independent tests were used to examine the fraction of apoptotic cells. Doxorubicin (DOX) was used as a positive control for cell death.
Kga108
Manufactured by Keygen Biotech
Sourced in China
The KGA108 is a compact desktop centrifuge designed for general laboratory applications. It features a maximum speed of 6,000 RPM and a maximum relative centrifugal force (RCF) of 3,683 x g. The centrifuge accommodates 8 x 15 mL tubes or 4 x 50 mL tubes in a fixed-angle rotor. The unit includes a digital display for speed and time settings.
Automatically generated - may contain errors
Lab products found in correlation
A00 1 1102, by Beckman Coulter (2 mentions)
Facscalibur flow cytometer, by BD (2 mentions)
Fc500 flow cytometry, by Beckman Coulter (1 mentions)
Flow cytometry, by BD (1 mentions)
Axio observer 5, by Zeiss (1 mentions)
Kga512, by Keygen Biotech (1 mentions)
In situ cell death detection kit, by Roche (1 mentions)
Facscalibur, by BD (1 mentions)
Hoechst 33342 staining, by Thermo Fisher Scientific (1 mentions)
Fluorescence microscope, by Olympus (1 mentions)
15 protocols using kga108
1
Apoptosis Induction in Glioma Cells
2
Annexin V-FITC Apoptosis Assay
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HBECs were harvested from 6-well plates and washed with PBS twice, centrifuged (800 rpm, 5 min), then mixed with 500 µL binding buffer. After resuspension, 10 µL propidium iodide (PI) and Annexin V-fluorescein isothiocyanate (FITC) were added, following the manufacturer’s instructions (KGA108, KeyGen, Nanjing, China). Finally, the HBECs were examined with the flow cytometer (A00-1-1102, Beckman, USA).
3
Annexin V and PI Apoptosis Assay
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Cells were stained with annexin V-FITC and PI (KGA108, KeyGEN) and evaluated for apoptosis by flow cytometry according to the manufacturer’s protocol. Briefly, 1 × 106 cells were washed twice with PBS and stained with 5 μl annexin V-FITC and 10 μl PI in 1 × binding buffer for 15 minutes at room temperature in the dark. Apoptotic cells were determined using a Beckman-Coulter Flow Cytometry FC500. Both early (annexin V-positive/PI-negative) and late (annexin V-positive/PI-positive) apoptotic cells were included when assessing cell death.
4
Probing miR-188-5p Regulation via Antagomir
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We applied miR-188-5p antagomir to probe the delicate regulation of miR-188-5p further. qRT-PCR results exhibited that miR-188-5p antagomir remarkably hindered miR-188-5p in vivo (Fig. 2A). Figure 2B indicated that MCAO/R-treated rats had a distinct infarction area in the brain's left hemisphere, while miR-188-5p antagomir reduced the brain-damaged area. Furthermore, miR-188-5p silencing signi cantly 2.12 Flow cytometry analysis
According to instructions (KGA108, keyGEN), PC12 cells were trypsinized and resuspended. 500 µL of 1 × binding buffer was supplemented, followed by 5 µL of Annexin V-FITC and Propidium Iodide. After incubation in the dark for 10 min, samples were subjected to ow cytometry (A00-1-1102, Beckman).
According to instructions (KGA108, keyGEN), PC12 cells were trypsinized and resuspended. 500 µL of 1 × binding buffer was supplemented, followed by 5 µL of Annexin V-FITC and Propidium Iodide. After incubation in the dark for 10 min, samples were subjected to ow cytometry (A00-1-1102, Beckman).
5
Apoptosis Measurement by Flow Cytometry
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Cells treated in each group were digested with trypsin without EDTA and collected. Then, PBS was used to wash the cells before centrifugation at 2000 rpm for 5 min. Approximately 5 × 105 cells were collected. Then, 500 μL of binding buffer was added to suspend the cells. Next, 5 μL of Annexin V-FITC (KGA108, KeyGen, China) and 5 μL of propidium iodide were thoroughly mixed. Finally, the cells were incubated at room temperature with the Annexin V-FITC/propidium iodide mixture in the dark for 15 min. Flow cytometry (A00-1-1102, Beckman, USA) was performed within 1 h of this treatment.
6
Annexin V-FITC/PI Apoptosis Assay
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The cells were collected after washing with PBS, following the instructions of Annexin V–FITC/propidium iodide (KGA108; KeyGen Biotech, Nanjing, Jiangsu, China). Subsequently, we added binding buffer suspension, Annexin V–FITC and propidium iodide. After mixing, the cells were incubated for 15 min. Apoptosis was detected by flow cytometry (Becton, Dickinson and Company, NJ, USA).
7
Quantifying Cell Death via Annexin V-FITC and PI
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Cell death was analyzed by ANXA5/annexin V-FITC-propidium iodide (PI) or PI staining. Cells were seeded at a density of 2 × 105 cells/well in 6-well plates. The next day, cells were incubated with the indicated treatments. For ANXA5-FITC-PI staining, cells were collected, washed with binding buffer, and then incubated in binding buffer with ANXA5-FITC and PI (KeyGEN BioTECH, KGA108) for 15 min in the dark. Dead cells were determined by flow cytometry. For PI staining, the cells were stained with PI for 30 min in an incubator of 5% CO2 at 37°C. The PI-positive cells were imaged by using a fluorescence microscope (ZEISS, Axio Observer 5).
8
Analyzing Cell Cycle and Apoptosis by Flow Cytometry
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Consistent with the previous description, cell cycle distribution, and apoptosis were detected by flow cytometry [24 (link), 36 (link), 37 (link)]. For cell cycle assay, after transfected with Importin-7 siRNAs, cells were digested and washed twice with 4 °C PBS and fixed with a mixture containing 2 ml cold 70% ethanol solution and 500 μl PBS at 4 °C overnight. After being washed with PBS, cells were prepared into a single cell suspension using a Propidium Iodide (PI)-containing cell cycle detection kit (KGA512, KeyGEN BioTECH, Nanjing, China) according to the instruction and left to stand in the dark for 30 min. Three independent tests for cell cycle distributions of each group were ultimately examined with flow cytometry.
For apoptosis assay, cells were treated according to the indicated conditions, then collected and washed twice using 4 °C PBS. An Annexin V-FITC/PI-containing apoptosis detection kit (KGA108, KeyGEN BioTECH, Nanjing, China) was used to stain the apoptotic cells according to the instruction. Flow cytometry was performed to determine the apoptotic rate from three independent experiments.
For apoptosis assay, cells were treated according to the indicated conditions, then collected and washed twice using 4 °C PBS. An Annexin V-FITC/PI-containing apoptosis detection kit (KGA108, KeyGEN BioTECH, Nanjing, China) was used to stain the apoptotic cells according to the instruction. Flow cytometry was performed to determine the apoptotic rate from three independent experiments.
9
Quantifying Apoptosis in Intervertebral Disc Cells
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TUNEL Assay (Terminal deoxynucleotidyl transferase dUTP nick end labeling) is a
common method for detecting apoptotic signaling cascades by labeling DNA
fragmentation. For TUNEL assay, NP cells were cultured in 12-well plates with
the density of 105 cells/well. After treatment with pramlintide, the
harvested NP cells were analyzed using in situ cell death detection kit
(12156792910, Roche, Branford, CT, USA) after fixing the cells in 4%
paraformaldehyde (pH 7.4). For cytometry analysis, the NP cells from each
treatment group were labeled with Annexin V-APC/7-AAD double staining, according
to the manufacturer’s instructions (KGA108, KeyGen Biotech, Nanjing, China).
Each sample was applied to a FACSCalibur Flow Cytometer (BD Biosciences, USA)
and fluorescence intensity of the apoptotic cells was detected.
common method for detecting apoptotic signaling cascades by labeling DNA
fragmentation. For TUNEL assay, NP cells were cultured in 12-well plates with
the density of 105 cells/well. After treatment with pramlintide, the
harvested NP cells were analyzed using in situ cell death detection kit
(12156792910, Roche, Branford, CT, USA) after fixing the cells in 4%
paraformaldehyde (pH 7.4). For cytometry analysis, the NP cells from each
treatment group were labeled with Annexin V-APC/7-AAD double staining, according
to the manufacturer’s instructions (KGA108, KeyGen Biotech, Nanjing, China).
Each sample was applied to a FACSCalibur Flow Cytometer (BD Biosciences, USA)
and fluorescence intensity of the apoptotic cells was detected.
10
Annexin V-FITC Apoptosis Assay
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An Annexin V-FITC assay [28 (link)] was used to quantify numbers of apoptotic cells by flow cytometry according to the manufacturer’s instructions (Nanjing Keygen Biotech, KGA108). Briefly, cells were collected following trypsinization, washed twice with ice-cold PBS, and resuspended in 300 μL of 1× binding buffer containing 5 μL Annexin V-FITC and 5 μL PI for 30 min at room temperature in the dark. All samples were analyzed on a FACSCalibur flow cytometer (BD Biosciences).
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