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KCL-ENG-301
⚠ FOR RESEARCH USE ONLY — Not for diagnostic or therapeutic procedures
KCL-ENG-301 is a stable, adherent HeLa cell line engineered to express full-length human Angiotensin-Converting Enzyme 2 (ACE2) fused with a C-terminal Red Fluorescent Protein (RFP) tag. The ACE2-RFP transgene is delivered via lentiviral or PiggyBac vector under a CMV promoter and maintained under Puromycin selection for stable integration. Greater than 95% of cells are RFP-positive by flow cytometry, with MFI at least 10-fold over the parental HeLa line.
ACE2 is the primary cellular entry receptor for SARS-CoV-2 and a key target in coronavirus research. The RFP tag enables direct live-cell visualization of ACE2 localization, membrane dynamics, and receptor trafficking without the need for fixation or antibody staining. Functional ACE2 enzymatic activity can be confirmed using a fluorogenic substrate assay (e.g., Mca-APK(Dnp)-OH). Cell line identity is confirmed by STR profiling against the HeLa reference.
All lots are quality tested prior to release for RFP expression, post-thaw viability, mycoplasma, sterility, and STR identity.
DMEM supplemented with 10% heat-inactivated FBS and 1% Pen/Strep. For initial recovery after thawing, culture without Puromycin for 48 hours. After the first passage, add Puromycin at 0.5 μg/mL, increasing to 1 to 2 μg/mL for routine maintenance. Omit Puromycin during experimental assays as needed. Subculture at 80 to 90% confluence using 0.25% Trypsin-EDTA; split 1:4 to 1:6, refeeding every 2 to 3 days.
All lots are quality tested prior to release. The following characterization data is generated for each production lot.
| Parameter | Result | Specification |
|---|---|---|
| ACE2-RFP expression | ✓ Positive | >95% RFP-positive by flow cytometry; MFI ≥10× parental HeLa |
| RFP fluorescence | ✓ Positive | Ex/Em 558/583 nm; detectable by PE/TRITC channel; cytoplasmic and membrane-associated signal |
| ACE2 protein (Western blot) | ✓ Confirmed | Anti-ACE2 antibody; expected band at fusion protein size |
| Post-thaw viability | ✓ Pass | >90% by Trypan Blue exclusion |
| Cell line identity (STR) | ✓ Confirmed | Matches HeLa reference STR profile |
| Transgene | ✓ Confirmed | Full-length human ACE2 (NP_001358344.1); C-terminal RFP fusion; CMV promoter |
| Test | Result | Method |
|---|---|---|
| Mycoplasma | ✗ Negative | PCR or luminescence assay |
| Sterility — Bacteria & Fungi | ✗ Negative | 14-day culture |
Certificates of Analysis with lot-specific QC data including flow cytometry histograms and STR profile are available upon request. Contact Kulture Scientific with your lot number.
KCL-ENG-301 is shipped as a cryopreserved vial on dry ice. These are adherent HeLa cells. Follow the steps below upon receipt.
Subculture at 80 to 90% confluence. Remove medium, rinse with PBS, add 0.25% Trypsin-EDTA for 2 to 3 minutes at 37°C. Neutralize with complete medium and split 1:4 to 1:6. Refeed every 2 to 3 days. Doubling time is approximately 24 hours. Maintain under Puromycin selection (1 to 2 μg/mL) to preserve stable ACE2-RFP expression. Full protocols are available on the Kulture Scientific protocol page.
KCL-ENG-301 is available as a cryopreserved vial containing at least 5×10&sup5; viable cells in cryoprotective medium.
| Product | Cat. No. | Format | Cell Number |
|---|---|---|---|
| Human ACE2 (RFP) HeLa Cell Line | KCL-ENG-301 | Cryopreserved Vial | ≥5×105 cells/vial |
- Storage — store in liquid nitrogen vapor phase (−150°C to −196°C); this is the only recommended storage condition for cryopreserved cells
- Do not store at −80°C or −20°C — neither temperature is suitable for long-term cell storage; ice crystal formation at these temperatures will irreversibly damage cell membranes and compromise viability
- Temperature sensitivity — minimize time outside cryogenic storage; cells are sensitive to temperature fluctuations
- Freeze-thaw cycles — repeated freeze-thaw cycles compromise viability and ACE2-RFP expression stability; thaw only the quantity needed for immediate use
- Stability — cells stored in liquid nitrogen are stable indefinitely when maintained under correct cryogenic conditions
- Shipping condition — cryopreserved vials are shipped on dry ice; inspect packaging upon receipt and transfer to liquid nitrogen storage immediately
- DMSO content — cryoprotective medium contains DMSO; wear nitrile gloves during thaw procedures and avoid skin contact
- Certificate of Analysis — lot-specific QC documentation including RFP flow cytometry data, viability, STR profile, and sterility results available upon request
For research purposes only. Not for use in diagnostic or therapeutic procedures. This product has not been tested or validated for clinical use. Not for direct administration into humans or animals.
This cell line should be handled under Biosafety Level 2 (BSL-2) conditions based on the following characteristics:
- Human-derived origin — HeLa cells are derived from human cervical adenocarcinoma tissue; all human-derived cell lines require BSL-2 containment under Universal Precautions
- HPV-18 sequences — HeLa cells are known to harbor integrated HPV-18 sequences; handle accordingly with BSL-2 precautions
- Lentiviral / PiggyBac transduction — the line was generated using a lentiviral or PiggyBac vector; the transgene does not contain viral replication genes and cannot produce infectious virus; standard BSL-2 containment applies
- ACE2 transgene — expresses human ACE2, the SARS-CoV-2 entry receptor; the transgene is non-infectious; however, this cell line may be used in co-culture with Spike-expressing or pseudovirus systems which may require additional biosafety considerations
Users must consult their institutional biosafety committee (IBC) prior to use, particularly if this line will be used in combination with SARS-CoV-2 pseudovirus or Spike-expressing systems.
- Human-derived material with HPV-18 — treat as potentially infectious; work exclusively in a certified biosafety cabinet; follow BSL-2 guidelines
- ACE2 expression — this line overexpresses the SARS-CoV-2 entry receptor; when used in pseudovirus entry assays, follow IBC guidance for the pseudovirus component
- DMSO (cryoprotectant) — skin-penetrating solvent; wear nitrile gloves at all times during thawing
- Waste disposal — autoclave all cell culture waste or treat with 10% bleach prior to disposal in accordance with local biohazardous waste regulations
⚠ FOR RESEARCH USE ONLY — Not for diagnostic or therapeutic procedures
KCL-ENG-301 is a stable, adherent HeLa cell line engineered to express full-length human Angiotensin-Converting Enzyme 2 (ACE2) fused with a C-terminal Red Fluorescent Protein (RFP) tag. The ACE2-RFP transgene is delivered via lentiviral or PiggyBac vector under a CMV promoter and maintained under Puromycin selection for stable integration. Greater than 95% of cells are RFP-positive by flow cytometry, with MFI at least 10-fold over the parental HeLa line.
ACE2 is the primary cellular entry receptor for SARS-CoV-2 and a key target in coronavirus research. The RFP tag enables direct live-cell visualization of ACE2 localization, membrane dynamics, and receptor trafficking without the need for fixation or antibody staining. Functional ACE2 enzymatic activity can be confirmed using a fluorogenic substrate assay (e.g., Mca-APK(Dnp)-OH). Cell line identity is confirmed by STR profiling against the HeLa reference.
All lots are quality tested prior to release for RFP expression, post-thaw viability, mycoplasma, sterility, and STR identity.
DMEM supplemented with 10% heat-inactivated FBS and 1% Pen/Strep. For initial recovery after thawing, culture without Puromycin for 48 hours. After the first passage, add Puromycin at 0.5 μg/mL, increasing to 1 to 2 μg/mL for routine maintenance. Omit Puromycin during experimental assays as needed. Subculture at 80 to 90% confluence using 0.25% Trypsin-EDTA; split 1:4 to 1:6, refeeding every 2 to 3 days.
All lots are quality tested prior to release. The following characterization data is generated for each production lot.
| Parameter | Result | Specification |
|---|---|---|
| ACE2-RFP expression | ✓ Positive | >95% RFP-positive by flow cytometry; MFI ≥10× parental HeLa |
| RFP fluorescence | ✓ Positive | Ex/Em 558/583 nm; detectable by PE/TRITC channel; cytoplasmic and membrane-associated signal |
| ACE2 protein (Western blot) | ✓ Confirmed | Anti-ACE2 antibody; expected band at fusion protein size |
| Post-thaw viability | ✓ Pass | >90% by Trypan Blue exclusion |
| Cell line identity (STR) | ✓ Confirmed | Matches HeLa reference STR profile |
| Transgene | ✓ Confirmed | Full-length human ACE2 (NP_001358344.1); C-terminal RFP fusion; CMV promoter |
| Test | Result | Method |
|---|---|---|
| Mycoplasma | ✗ Negative | PCR or luminescence assay |
| Sterility — Bacteria & Fungi | ✗ Negative | 14-day culture |
Certificates of Analysis with lot-specific QC data including flow cytometry histograms and STR profile are available upon request. Contact Kulture Scientific with your lot number.
KCL-ENG-301 is shipped as a cryopreserved vial on dry ice. These are adherent HeLa cells. Follow the steps below upon receipt.
Subculture at 80 to 90% confluence. Remove medium, rinse with PBS, add 0.25% Trypsin-EDTA for 2 to 3 minutes at 37°C. Neutralize with complete medium and split 1:4 to 1:6. Refeed every 2 to 3 days. Doubling time is approximately 24 hours. Maintain under Puromycin selection (1 to 2 μg/mL) to preserve stable ACE2-RFP expression. Full protocols are available on the Kulture Scientific protocol page.
KCL-ENG-301 is available as a cryopreserved vial containing at least 5×10&sup5; viable cells in cryoprotective medium.
| Product | Cat. No. | Format | Cell Number |
|---|---|---|---|
| Human ACE2 (RFP) HeLa Cell Line | KCL-ENG-301 | Cryopreserved Vial | ≥5×105 cells/vial |
- Storage — store in liquid nitrogen vapor phase (−150°C to −196°C); this is the only recommended storage condition for cryopreserved cells
- Do not store at −80°C or −20°C — neither temperature is suitable for long-term cell storage; ice crystal formation at these temperatures will irreversibly damage cell membranes and compromise viability
- Temperature sensitivity — minimize time outside cryogenic storage; cells are sensitive to temperature fluctuations
- Freeze-thaw cycles — repeated freeze-thaw cycles compromise viability and ACE2-RFP expression stability; thaw only the quantity needed for immediate use
- Stability — cells stored in liquid nitrogen are stable indefinitely when maintained under correct cryogenic conditions
- Shipping condition — cryopreserved vials are shipped on dry ice; inspect packaging upon receipt and transfer to liquid nitrogen storage immediately
- DMSO content — cryoprotective medium contains DMSO; wear nitrile gloves during thaw procedures and avoid skin contact
- Certificate of Analysis — lot-specific QC documentation including RFP flow cytometry data, viability, STR profile, and sterility results available upon request
For research purposes only. Not for use in diagnostic or therapeutic procedures. This product has not been tested or validated for clinical use. Not for direct administration into humans or animals.
This cell line should be handled under Biosafety Level 2 (BSL-2) conditions based on the following characteristics:
- Human-derived origin — HeLa cells are derived from human cervical adenocarcinoma tissue; all human-derived cell lines require BSL-2 containment under Universal Precautions
- HPV-18 sequences — HeLa cells are known to harbor integrated HPV-18 sequences; handle accordingly with BSL-2 precautions
- Lentiviral / PiggyBac transduction — the line was generated using a lentiviral or PiggyBac vector; the transgene does not contain viral replication genes and cannot produce infectious virus; standard BSL-2 containment applies
- ACE2 transgene — expresses human ACE2, the SARS-CoV-2 entry receptor; the transgene is non-infectious; however, this cell line may be used in co-culture with Spike-expressing or pseudovirus systems which may require additional biosafety considerations
Users must consult their institutional biosafety committee (IBC) prior to use, particularly if this line will be used in combination with SARS-CoV-2 pseudovirus or Spike-expressing systems.
- Human-derived material with HPV-18 — treat as potentially infectious; work exclusively in a certified biosafety cabinet; follow BSL-2 guidelines
- ACE2 expression — this line overexpresses the SARS-CoV-2 entry receptor; when used in pseudovirus entry assays, follow IBC guidance for the pseudovirus component
- DMSO (cryoprotectant) — skin-penetrating solvent; wear nitrile gloves at all times during thawing
- Waste disposal — autoclave all cell culture waste or treat with 10% bleach prior to disposal in accordance with local biohazardous waste regulations
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