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Internal Reference:
KCL-ENG-202
⚠ FOR RESEARCH USE ONLY — Not for diagnostic or therapeutic procedures
KCL-ENG-202 is a stable, adherent HeLa cell line engineered to constitutively express the full-length SARS-CoV-2 Spike (S) glycoprotein under a CMV promoter. The Spike protein carries a C-terminal enhanced GFP (eGFP) tag for direct fluorescence visualization and flow cytometry detection without requiring anti-Spike antibody staining in every assay. The line is maintained under Puromycin selection to ensure stable transgene retention.
Greater than 85% of cells co-express Spike and eGFP as confirmed by flow cytometry. Surface Spike expression is validated by anti-Spike antibody staining (92% positive; MFI greater than 10,000), and functional receptor interaction is confirmed by dose-dependent binding of recombinant human ACE2-Fc. Western blot shows the expected full-length band at approximately 180 kDa. Use within 10 passages post-thaw to ensure consistent stable expression.
This cell line does not produce infectious SARS-CoV-2 virus. It expresses Spike protein only and is suitable for BSL-2 work. All lots are quality tested prior to release for Spike/GFP co-expression, viability, mycoplasma, and sterility.
DMEM supplemented with 10% heat-inactivated FBS and 1% Pen/Strep. Maintain under Puromycin selection at 1 μg/mL for routine culture. Omit Puromycin during experimental assays as needed. Subculture at 80 to 90% confluence using 0.25% Trypsin-EDTA; split 1:4 to 1:6 every 2 to 3 days.
All lots are quality tested prior to release. Expression validation is performed at passage 4. The following data is representative of lot release testing.
| Parameter | Result | Specification |
|---|---|---|
| SARS-CoV-2 Spike surface expression | ✓ Positive | 92% by flow cytometry (anti-Spike clone 1A9); MFI >10,000 |
| eGFP fluorescence | ✓ Positive | 89% by flow cytometry (GFP channel); C-terminal co-localized with Spike |
| Spike / eGFP co-expression | ✓ Confirmed | >85% double-positive of live cells |
| Spike protein (Western blot) | ✓ Confirmed | Band at ~180 kDa (full-length); anti-S1/RBD antibody; reducing conditions |
| ACE2 binding (functional) | ✓ Positive | Dose-dependent binding of recombinant human ACE2-Fc; ≥1-log shift over parental HeLa |
| Post-thaw viability | ✓ Pass | >90% by Trypan Blue exclusion |
| Passage number at freeze | ✓ Confirmed | Passage 1 |
| Test | Result | Method |
|---|---|---|
| Mycoplasma | ✗ Negative | PCR (EP 2.6.7) |
| Sterility — Bacteria | ✗ Negative | 14-day culture |
| Sterility — Fungi & Yeast | ✗ Negative | 14-day culture |
Certificates of Analysis with lot-specific QC data including flow cytometry data for Spike and GFP co-expression are available upon request. Contact Kulture Scientific with your lot number.
KCL-ENG-202 is shipped as a cryopreserved vial on dry ice. These are adherent HeLa cells. Follow the steps below upon receipt. Process within 48 hours of delivery.
Subculture at 80 to 90% confluence using 0.25% Trypsin-EDTA. Split 1:4 to 1:6 every 2 to 3 days. Maintain under Puromycin selection at 1 μg/mL. Verify Spike and eGFP co-expression by flow cytometry periodically, particularly before critical experiments. Re-establish from frozen stock after passage 10. Full protocols available on the Kulture Scientific protocol page.
KCL-ENG-202 is available as a cryopreserved vial containing at least 5×10&sup5; viable cells in cryoprotective medium.
| Product | Cat. No. | Format | Cell Number |
|---|---|---|---|
| COVID-19 Spike (S) Protein / HeLa Cell Line | KCL-ENG-202 | Cryopreserved Vial | ≥5×105 cells/vial |
- Storage — store in liquid nitrogen vapor phase (−150°C to −196°C); cells are stable indefinitely under correct cryogenic conditions
- 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 Spike/eGFP co-expression stability; thaw only the quantity needed for immediate use
- Shipping condition — cryopreserved vials are shipped on dry ice; process within 48 hours of delivery and transfer to liquid nitrogen storage immediately upon receipt
- Post-thaw passage limit — use within 10 passages post-thaw; re-establish from frozen stock after passage 10 to maintain stable expression
- DMSO content — cryoprotective medium contains DMSO; wear nitrile gloves during thaw procedures and avoid skin contact
- Certificate of Analysis — lot-specific QC documentation including Spike and GFP flow cytometry data and viability results available upon request
For research purposes only. Not for use in diagnostic or therapeutic procedures. Not for in vivo use without further validation. 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. This product does not contain or produce infectious SARS-CoV-2 virus. BSL-2 is required based on the following:
- Human-derived origin — HeLa cells are derived from human cervical adenocarcinoma tissue; all human-derived cell lines require BSL-2 containment under Universal Precautions
- SARS-CoV-2 Spike transgene — expresses the viral Spike glycoprotein; does not contain the full SARS-CoV-2 genome and cannot produce infectious virus; not a select agent
- Recombinant cell line — stable lentiviral or plasmid integration; standard BSL-2 containment for recombinant human cell lines applies
Users must consult their institutional biosafety committee (IBC) prior to use. Individual institutions may impose additional requirements based on local policy regarding SARS-CoV-2-related research materials.
- Human-derived material — treat as potentially infectious; work in a certified biosafety cabinet; follow BSL-2 guidelines at all times
- Spike protein expression — this cell line expresses SARS-CoV-2 Spike glycoprotein on the cell surface; it cannot produce infectious virus but should be treated with appropriate containment
- Spill response — cover with 10% bleach, allow 20 minutes contact time, absorb, and decontaminate the surface; document all spill incidents per institutional protocol
- 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-202 is a stable, adherent HeLa cell line engineered to constitutively express the full-length SARS-CoV-2 Spike (S) glycoprotein under a CMV promoter. The Spike protein carries a C-terminal enhanced GFP (eGFP) tag for direct fluorescence visualization and flow cytometry detection without requiring anti-Spike antibody staining in every assay. The line is maintained under Puromycin selection to ensure stable transgene retention.
Greater than 85% of cells co-express Spike and eGFP as confirmed by flow cytometry. Surface Spike expression is validated by anti-Spike antibody staining (92% positive; MFI greater than 10,000), and functional receptor interaction is confirmed by dose-dependent binding of recombinant human ACE2-Fc. Western blot shows the expected full-length band at approximately 180 kDa. Use within 10 passages post-thaw to ensure consistent stable expression.
This cell line does not produce infectious SARS-CoV-2 virus. It expresses Spike protein only and is suitable for BSL-2 work. All lots are quality tested prior to release for Spike/GFP co-expression, viability, mycoplasma, and sterility.
DMEM supplemented with 10% heat-inactivated FBS and 1% Pen/Strep. Maintain under Puromycin selection at 1 μg/mL for routine culture. Omit Puromycin during experimental assays as needed. Subculture at 80 to 90% confluence using 0.25% Trypsin-EDTA; split 1:4 to 1:6 every 2 to 3 days.
All lots are quality tested prior to release. Expression validation is performed at passage 4. The following data is representative of lot release testing.
| Parameter | Result | Specification |
|---|---|---|
| SARS-CoV-2 Spike surface expression | ✓ Positive | 92% by flow cytometry (anti-Spike clone 1A9); MFI >10,000 |
| eGFP fluorescence | ✓ Positive | 89% by flow cytometry (GFP channel); C-terminal co-localized with Spike |
| Spike / eGFP co-expression | ✓ Confirmed | >85% double-positive of live cells |
| Spike protein (Western blot) | ✓ Confirmed | Band at ~180 kDa (full-length); anti-S1/RBD antibody; reducing conditions |
| ACE2 binding (functional) | ✓ Positive | Dose-dependent binding of recombinant human ACE2-Fc; ≥1-log shift over parental HeLa |
| Post-thaw viability | ✓ Pass | >90% by Trypan Blue exclusion |
| Passage number at freeze | ✓ Confirmed | Passage 1 |
| Test | Result | Method |
|---|---|---|
| Mycoplasma | ✗ Negative | PCR (EP 2.6.7) |
| Sterility — Bacteria | ✗ Negative | 14-day culture |
| Sterility — Fungi & Yeast | ✗ Negative | 14-day culture |
Certificates of Analysis with lot-specific QC data including flow cytometry data for Spike and GFP co-expression are available upon request. Contact Kulture Scientific with your lot number.
KCL-ENG-202 is shipped as a cryopreserved vial on dry ice. These are adherent HeLa cells. Follow the steps below upon receipt. Process within 48 hours of delivery.
Subculture at 80 to 90% confluence using 0.25% Trypsin-EDTA. Split 1:4 to 1:6 every 2 to 3 days. Maintain under Puromycin selection at 1 μg/mL. Verify Spike and eGFP co-expression by flow cytometry periodically, particularly before critical experiments. Re-establish from frozen stock after passage 10. Full protocols available on the Kulture Scientific protocol page.
KCL-ENG-202 is available as a cryopreserved vial containing at least 5×10&sup5; viable cells in cryoprotective medium.
| Product | Cat. No. | Format | Cell Number |
|---|---|---|---|
| COVID-19 Spike (S) Protein / HeLa Cell Line | KCL-ENG-202 | Cryopreserved Vial | ≥5×105 cells/vial |
- Storage — store in liquid nitrogen vapor phase (−150°C to −196°C); cells are stable indefinitely under correct cryogenic conditions
- 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 Spike/eGFP co-expression stability; thaw only the quantity needed for immediate use
- Shipping condition — cryopreserved vials are shipped on dry ice; process within 48 hours of delivery and transfer to liquid nitrogen storage immediately upon receipt
- Post-thaw passage limit — use within 10 passages post-thaw; re-establish from frozen stock after passage 10 to maintain stable expression
- DMSO content — cryoprotective medium contains DMSO; wear nitrile gloves during thaw procedures and avoid skin contact
- Certificate of Analysis — lot-specific QC documentation including Spike and GFP flow cytometry data and viability results available upon request
For research purposes only. Not for use in diagnostic or therapeutic procedures. Not for in vivo use without further validation. 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. This product does not contain or produce infectious SARS-CoV-2 virus. BSL-2 is required based on the following:
- Human-derived origin — HeLa cells are derived from human cervical adenocarcinoma tissue; all human-derived cell lines require BSL-2 containment under Universal Precautions
- SARS-CoV-2 Spike transgene — expresses the viral Spike glycoprotein; does not contain the full SARS-CoV-2 genome and cannot produce infectious virus; not a select agent
- Recombinant cell line — stable lentiviral or plasmid integration; standard BSL-2 containment for recombinant human cell lines applies
Users must consult their institutional biosafety committee (IBC) prior to use. Individual institutions may impose additional requirements based on local policy regarding SARS-CoV-2-related research materials.
- Human-derived material — treat as potentially infectious; work in a certified biosafety cabinet; follow BSL-2 guidelines at all times
- Spike protein expression — this cell line expresses SARS-CoV-2 Spike glycoprotein on the cell surface; it cannot produce infectious virus but should be treated with appropriate containment
- Spill response — cover with 10% bleach, allow 20 minutes contact time, absorb, and decontaminate the surface; document all spill incidents per institutional protocol
- 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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