Normal mouse liver AML‐12 cells, human normal liver cells (LO2) and HEK‐293T cells were inoculated in 24‐well culture plates (1 × 104 cells per well) supplied with 0.5 mL DMEM (containing 10% FBS) for 24 h before transfection. Lipo8k was included as a positive control and the different groups were set to a medium containing no serum and 10% serum. Both lipo8K‐mRNA and LNP@GFP‐mRNA containing 1 µg GFP‐mRNA were added to 24‐well plates for 24 h incubation at 37 °C. Finally, inverted fluorescence microscopy (Nikon, Japan) and FACS (ACEA Bioscience) were used to evaluate the transfection effect. For the transfection stability test, 293T cells were transfected with LNP@GFP‐mRNA, at all sampled time points within one month.
Fluorescence activated cell sorting (facs)
Manufactured by Agilent Technologies
Sourced in United States
FACS (Fluorescence-Activated Cell Sorting) is a specialized type of flow cytometry that allows for the separation and isolation of specific cells from a heterogeneous population. It uses the principles of light scattering, light excitation, and electronic detection to analyze and physically separate cells based on their specific light-scattering and fluorescent characteristics.
Automatically generated - may contain errors
Lab products found in correlation
Edu alexa fluor 647 kit, by Keygen Biotech (3 mentions)
Annexin 5 propidium iodide, by Keygen Biotech (2 mentions)
Binding buffer, by Keygen Biotech (2 mentions)
Inverted fluorescence microscopy, by Nikon (1 mentions)
Accuri c6, by BD (1 mentions)
Fitc conjugated anti human cd45 ab, by BD (1 mentions)
Annexin 5 apc 7 aad, by Keygen Biotech (1 mentions)
7 protocols using fluorescence activated cell sorting (facs)
1
Evaluating mRNA Transfection Efficiency
2
Quantifying Cell Apoptosis with Annexin V-PI Staining
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell apoptosis was analyzed by staining with Annexin V/propidium iodide (PI) (Nanjing KeyGen Biotech Co., Ltd.). Briefly, ARPE19 cells were treated with HG for 24 h. The cells were washed twice with PBS and resuspended in 500 µl 1X binding buffer (Nanjing KeyGen Biotech Co., Ltd.), followed by incubation at 25°C for 15 min in the dark. Fluorescence of PI and Annexin V was monitored by FACS (ACEA Bio-science, Inc.) at 525 and 630 nm, respectively. The data was analyzed using BD Accuri C6 software (BD Biosciences).
3
Cell Proliferation Analysis by EdU Flow
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were stained with EdU Alexa Fluor 647 kit (#KGA334-50, Keygen; Jiangsu, China) according to the manufacturer’s protocol. The stained cells were analyzed by flow cytometry (FACS; ACEA Biosciences; San Diego, CA, USA).
4
Cell Cycle Analysis by EdU Staining
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were stained with EdU Alexa Fluor 647 kit (Keygen; Jiangsu, China) according to the manufacturer's protocol. The stained cells were analyzed by FACS (ACEA Biosciences).
5
PLAGL2 Overexpression and Apoptosis Resistance
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HCC cells were plated in six‐well plates (4 × 105 cells/well). After 24 h incubation, the cells were transfected with plasmids for 24 h or treated with SeS2 (0, 5, 10, and 20 μM) for 48 h. For PLAGL2 plasmid transfection, the vector plasmids were transfected and used as control groups. The cells were then treated with the apoptosis inducers CCCP (25 μM) and etoposide (40 μM) for 24 h in vector‐transfected control and PLAGL2‐overexpressing Bel‐7402 and Bel‐7404 cells to investigate the effect of PLAGL2 on apoptosis resistance. The cells were then harvested, and 5 μL Annexin V‐PE and 5 μL 7AAD staining solution were added to stain the cells. Apoptosis was detected using FACS (ACEA, Hangzhou, China).
6
Evaluating Leukemia Cell Proliferation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were co-cultured with stromal cells for 48 h, harvested, washed, xed, permeabilized, and incubated with FITC-conjugated PHA-E for 2 h at 37°C. MDS/AML malignant clonal cells were distinguished from co-cultured cells using FITC-conjugated anti-human CD45 Ab (BD Biosciences; Franklin Lakes, NJ, USA).
For proliferation analysis, cells were stained with EdU Alexa Fluor 647 kit (Keygen; Jiangsu, China) as per manufacturer's protocol, and analyzed by FACS (ACEA Biosciences; San Diego, CA, USA).
For proliferation analysis, cells were stained with EdU Alexa Fluor 647 kit (Keygen; Jiangsu, China) as per manufacturer's protocol, and analyzed by FACS (ACEA Biosciences; San Diego, CA, USA).
7
Apoptosis Quantification by Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The apoptotic incidence was detected by Annexin V/propidium iodide (PI), Annexin V-APC/7-AAD and binding buffer (KeyGEN Biotech). Neuro 2A cells were treated with various treatments. Cells were then collected and re-suspended in 500μl 1xbinding buffer and then incubated for 15 minutes without light exposure. Apoptotic cells, including early apoptotic (Annexin V + /PI -) and late apoptotic/necrotic (Annexin V + /PI + ) cells, were counted using FACS (ACEA Biosciences, CA, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!