SiHa/EGFP_LC3
Cervical cancer cell line SiHa stably expressing EGFP-LC3 for use as an autophagy sensor.
Contributors
| Inventor | Institute |
|---|---|
| T.R. Santhosh Kumar | Rajiv Gandhi Centre for Biotechnology |
| Cat. #: | 161144 |
|---|---|
| Cancer types: | Gynaecologic cancer |
| Research Fields: | Cancer;Disease (not cancer);Metabolism |
| Organism: | Human |
| Tissue: | Cervix |
| Gender: | Female |
| Model: | Transgenic |
| Morphology: | Epithelial |
| Growth properties: | Adherent |
| Cancer Types In Detail: | Cervical |
| Primary citation: | Lupitha SS, et.al. PMID: 32109534. |
| Product description: | MAP1LC3B (Microtubule associated protein 1 Light Chain 3 ?) is a protein involved in the autophagic process. Under normal physiological conditions the protein is cytosolic and during autophagy it undergoes lipidation to involve in autophagosome elongation. This process can be visualized as LC3B punctae formation. The cell line stably express MAP1LC3B tagged with EGFP and allow easy identification and quantitation of cells with autophagy response using a wide variety of fluorescent imaging platforms including high content imaging. The cell line is an ideal cell tool for autophagy and apoptosis research and support long time imaging of autophagy in live cells. |
|---|---|
| Gender: | Female |
| CRISPR: | No |
| Conditional: | No |
| Production details: | The product is a human cervical carcinoma cell line SiHa, transfected to stably express the protein MAP1LC3B tagged with EGFP. The EGFP-tag enables easy identification and affinity purification of the recombinant protein. The cell line can be utilized for studying the autophagy process and to screen of potential candidate drugs that induce autophagy. Stably expressing single cell clones were propagated. The expression and functionality of exogenous EGFP-LC3-B was confirmed through fluorescenc… |
| Parental cell line: | SiHa |
| disease: | Cancer |
| Initial handling information: | Resuscitation: Rapidly thaw the frozen ampoule in a water bath at 37?°C for 1-2 minutes. Transfer the contents to a centrifuge tube and slowly add 5-10ml of pre-warmed growth media. Remove a sample for counting. Centrifuge at 100-150g for 2-3 minutes to pellet cells (a centrifugation step to remove the cryoprotectant is essential. Seed at the recommend density, e.g. greater than 10,000 cells per cm2. |
|---|---|
| Storage conditions: | Liquid Nitrogen |
| Shipping conditions: | Dry Ice |
| Growth medium: | DMEM with 2mM L-Glutamine and 10% FBS (Heat inactivated), or EMEM with 2mM L-Glutamine and 10% Fetal Bovine Serum |
| Subculture routine: | Subculture non-confluent cultures using trypsin/EDTA and 1:3 to 1:4, i.e. seeding at 1-4 x10,000 cells/cm? . |
| Temperature: | 37° C |
| Atmosphere: | 5% CO2 |
| Storage medium: | Growth medium + 10% DMSO |
| References: |
Lupitha SS, et.al. PMID: 32109534. |
|---|
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