Green fluorescent protein (GFP)-expressing lentivirus and TF-shRNA lentivirus were purchased commercially (Genechem, Shanghai, China). Briefly, a stem-loop structure oligonucleotide containing the TF target sequence and a GFP reporter were cloned under the control of the human U6 promoter in lentiviral vectors. The sequence of the TF-shRNA is 5′-GCGCUUCAGGCACUACAAA-3′. The transfection of A549 cells was performed according to the manufacturer’s protocol. Briefly, a total of 5 × 104 A549 cells were seeded into a 24-well plate. After cell adherence, 1 × 106 IU of lentivirus was added to the culture medium, at a final Multiplicity of infection (MOI) of 20. The lentivirus-containing medium was maintained for 24 h before being replaced by fresh culture medium. The transfected cells were cultured normally and enriched by fluorescence-activated cell sorting (FACS). The efficiency of transfection was evaluated by real-time Polymerase chain reaction (rt-PCR) and Western blot analysis.
Gfp expressing lentivirus
Manufactured by Genechem
The GFP-expressing lentivirus is a laboratory tool designed to produce recombinant lentiviral particles that express the green fluorescent protein (GFP) gene. This tool is commonly used in research applications to label and track cells, monitor gene expression, and study cellular processes.
Automatically generated - may contain errors
2 protocols using gfp expressing lentivirus
1
Lentivirus-mediated Knockdown of Transcription Factor
2
Immune Cell Culture and Activation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A20 murine lymphoma cells (ATCC TIB-208) were cultured in RPMI-1640 medium (Sigma-Aldrich, Shanghai, China) supplied with 10% FBS. EL4 murine lymphoma cells (ATCC TIB-39) were cultured in DMEM medium (Thermo Fisher Scientific, Waltham, MA) supplied with 10% FBS. Jurkat cells (ATCC TIB-152) were cultured in RPMI-1640 medium containing 10% FBS. A20 and EL4 cells, stably expressing green fluorescence protein (GFP), were established by transducing cells with GFPexpressing lentivirus (Genechem, Shanghai, China). T cells were isolated from spleen cells using a Mouse CD90.2 Positive Selection Kit II from STEMCELL Technologies (Shanghai, China). Purities of isolated T cells were greater than 90%. Isolated mouse T cells were cultured at a density of 1 × 10 6 /mL in RPMI-1640 medium containing 10% FBS, 3 μg/mL anti-mouse CD3 (145-2C11) and 3 μg/mL anti-mouse CD28 (37.51) (BioLegend, San Diego, CA). Anti-human CD3/CD28 (10 971, STEMCELL Technologies) was used to stimulate 1 × 10 6 Jurkat cells. Mixed lymphocyte reaction (MLR) was performed by co-culturing lymphocytes isolated from C57BL/6 and BALB/c mice at a density of 2 × 10 6 /mL in RPMI-1640 medium containing 10% FBS.
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!