HK EGFP-alpha-tubulin/H2B-mCherry Cells
$540.00*
Prices excl. VAT plus shipping costs This cell line is licensed to CLS by a university or institute. The sale of this item requires the conclusion of a Material Transfer Agreement (MTA). Please get in touch with us for further information.
General information
Description | This HeLa reporter cell line stably expresses a red chromatin marker (core histone 2B fused to monomeric Cherry; H2B-mCherry; transfection plasmid: pH2B-mCherry-IRES-neo3) and a marker for microtubules (monomeric enhanced GFP-?-tubulin; transfection plasmid: pmEGFP-a-tubulin-IRES-puro2b). This clonal stable cell line was generated by transfection of a circular plasmid followed by drug resistance selection. |
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Organism | Human |
Tissue | Cervix |
Disease | Carcinoma |
Synonyms | HeLa Kyoto EGFP-a-tubulin/H2B-mCherry, HeLa H2B-mRFP and mEGFP-alpha-tubulin |
Characteristics
Age | 30 years |
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Gender | Female |
Ethnicity | African American |
Morphology | Epithelial-like cells with mosaic stone shape |
Growth properties | Monolayer, adherent |
Identifiers / Biosafety / Citation
Citation | HK EGFP-alpha-tubulin/H2B-mCherry (Cytion catalog number 300670) |
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Biosafety level | 1 |
Depositor | Dr. J. Ellenberg, EMBL Heidelberg |
Expression / Mutation
Protein expression | EGFP-alpha-tubulin, H2B-mCherry: Location/Gene: 1..589 / Pcmv, 652..1029 H2B, 1042..1752 / mCherry, 2983..3777 / KanR/NeoR |
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Viruses | Negative for HIV, HBV and HCV. |
Products | CMV Promotor, Histone H2B, Neomycin, Phosphotransferase |
Handling
Culture Medium | DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 1.5 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a) |
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Medium supplements | Supplement the medium with 10% FBS |
Passaging solution | Accutase |
Doubling time | 24 hours |
Subculturing | Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium. |
Split ratio | A ratio of 1:3 is recommended |
Seeding density | 1 x 10^4 cells/cm^2 |
Fluid renewal | 2 to 3 times per week |
Freezing recovery | After thawing, plate the cells at 5 x 10^4 cells/cm^2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours. |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures |
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Quality control / Genetic profile / HLA
Sterility | Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. |
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