| Field | Specification |
|---|---|
| Product Format | Frozen |
| Product Type | |
| Shipping | |
| Storage |
Overview
MicroRNAs (miRNAs) comprise a class of small non-coding RNAs that regulate the stability and/or translatability of most protein-coding transcripts. They guide AGO proteins within the RNA-induced silencing complex (RISC) and plays a part to degrade or repress the translation of their target mRNAs.
Key elements and design rationale
- Model identity: 3xFLAG-AGO2 Stable Expressing HEK293T Cell Line is supplied as an engineered cell line derived from Human kidney.
- Growth properties: Adherent, epithelial
- Growth conditions: PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) are recommended for optimal cell culture. Dulbecco's Modified Eagle Medium (DMEM), High Glucose (TM500) + 5% FBS + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂.
- Product format: Frozen, BSL-2
This cell-based model is generally used in kidney biology, phenotype comparison, and assay development studies. Donor/background information is available for contextual interpretation.
Biological background
3xFLAG-AGO2 stable expressing HEK293T cells can allow you to investigate the susceptibility of AGO2-loaded miRNA to different classes of nucleases and can be used in complement to in vivo studies to identify new miRNA decay factors. Donor/background information provided for this product: Embryo.
Research relevance and current trends
- Engineered cell lines are widely used for reporter-based readouts, perturbation studies, and assay optimization in reproducible culture systems.
- Reporter or transgene-bearing models are often compared with matched parental or control cells to interpret signal changes in context.
- Expression trends are typically evaluated alongside passage number, selection pressure, and baseline growth behavior.
Common research applications
- Routine expansion and maintenance of a defined cell model for downstream in vitro experiments.
- Phenotype, signaling, or marker-expression studies performed under standardized culture conditions.
- Cell-based assay development in which passage number, growth surface, and medium composition are tracked as experimental variables.
Changes in morphology, growth rate, viability, or reporter signal are typically interpreted together with passage history, culture matrix, and the specified growth conditions for the model.
Notes for experimental interpretation
- Morphology, doubling behavior, and reporter or marker output can shift with passage number, substrate choice, and medium composition; these variables should be recorded alongside experimental readouts.
- Matched controls such as parental cells, untreated cultures, or parallel cultures maintained under identical conditions help distinguish background effects from biology of interest.
Culture and product details
- Growth Conditions: PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) are recommended for optimal cell culture. Dulbecco's Modified Eagle Medium (DMEM), High Glucose (TM500) + 5% FBS + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂.
- Seeding Density (cells/cm²): 20,000 - 40,000
- Thaw cells quickly in a 37°C water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination.
- Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions.
- Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media. Centrifuge cells at 125xg for 5-7 minutes.
- Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask.
- Incubate the cells at the recommended conditions.
- Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel.
- Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37°C, for several minutes to facilitate detachment.
- Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel.
- Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type.
- Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired.
- Incubate the cells at the recommended conditions.
How should I handle live cells once I receive them?
https://www.abmgood.com/immortalized-cells-documents.html
Following these guidelines will help ensure optimal cell viability and performance.
Why are these cells classified as biosafety level II?
What is your warranty or return policy?
Please refer to the following link for full information:
https://www.abmgood.com/terms
For additional questions, our Order team is happy to assist and can be reached at order@abmgood.com.
How many times can cells divide?
Primary cells have a limited lifespan and will undergo a finite number of population doublings before entering senescence. The exact number varies by cell type and culture conditions.
Immortalized cell lines are capable of extended or indefinite proliferation under proper culture conditions, although growth characteristics may vary between lines.
Do I need Applied Cell Extracellular Matrix (G422) if I am using PriCoat™ flasks?
Cell line sourcing and selection (species, tissue, and disease model matching) · Stable cell line engineering (overexpression, knockdown, knockout via CRISPR/Cas9, shRNA, sgRNA) · Reporter gene integration (GFP, RFP, luciferase, fluorescent/bioluminescent constructs) · Genome editing and knockin (point mutations, tagged endogenous proteins, conditional alleles) · Inducible expression systems (Tet-On/Off and regulatable constructs) · Drug resistance marker selection (puromycin, G418, hygromycin, and others) · Custom growth and media optimisation for specific assay requirements · Scale-up production for high-throughput screening campaigns · Authentication and QC services (STR profiling, mycoplasma testing, viability assessment). Talk to a Scientist or contact support@biohippo.com.
Elbarbary, R. A., & Maquat, L. E. (2019). Evaluating the susceptibility of AGO2-loaded microRNAs to degradation by nucleases in vitro. Methods (San Diego, Calif.), 152, 18–22. https://doi.org/10.1016/j.ymeth.2018.05.010
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