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KCL-ENG-302
⚠ FOR RESEARCH USE ONLY — Not for diagnostic or therapeutic procedures
KCL-ENG-302 is a stable HEK293T cell line constitutively expressing full-length human Angiotensin-Converting Enzyme 2 (ACE2) fused with a C-terminal Red Fluorescent Protein (RFP) tag. The ACE2-RFP transgene is stably integrated under a CMV promoter and maintained under Puromycin or G418 (Neomycin) selection — confirm the selection marker for your specific lot with the Certificate of Analysis.
Greater than 90% of cells are RFP-positive by flow cytometry. ACE2 mRNA expression is more than 100-fold over parental HEK293T cells by RT-qPCR. Functional ACE2 enzymatic activity is confirmed by fluorogenic peptide substrate assay (signal-to-background ratio of at least 5:1). Stable single-copy genomic integration is confirmed by PCR or Southern blot. Cell line identity is confirmed by STR profiling against the ATCC CRL-3216 HEK293T reference.
HEK293T is a high-transfection-efficiency derivative of HEK293 widely used in viral pseudotype and protein expression systems. The combination of high ACE2 expression, RFP reporter, and the HEK293T background makes this line well suited for SARS-CoV-2 pseudovirus entry assays, live-cell ACE2 receptor imaging, and drug screening targeting ACE2 activity or Spike/ACE2 interaction. Note: this cell line does not express TMPRSS2; for Spike-primed viral entry, co-express TMPRSS2 or supplement with exogenous trypsin.
DMEM supplemented with 10% heat-inactivated FBS and 1% Pen/Strep. Maintain under Puromycin (1 μg/mL) or G418 (500 μg/mL) selection as appropriate for your lot. Do not maintain cells under selection antibiotic for more than 4 weeks without re-qualifying expression. Subculture at 80 to 90% confluence using a cell detachment solution; split 1:4 to 1:6 every 2 to 3 days. Do not vortex cells.
All lots are quality tested prior to release. The following characterization data is generated for each production lot.
| Parameter | Result | Specification |
|---|---|---|
| ACE2-RFP expression (flow cytometry) | ✓ Positive | ≥90% RFP-positive cells; Ex/Em 558/583 nm |
| ACE2 surface expression | ✓ Positive | High surface expression by immunofluorescence or flow cytometry (anti-ACE2 antibody) |
| ACE2 mRNA expression (RT-qPCR) | ✓ Confirmed | >100-fold over parental HEK293T |
| ACE2 enzymatic activity | ✓ Active | Fluorogenic peptide assay (Mca-APK(Dnp)-OH); signal/background ≥5:1 |
| Genomic integration | ✓ Confirmed | Stable single-copy integration by PCR or Southern blot |
| Post-thaw viability | ✓ Pass | ≥70% by Trypan Blue exclusion |
| Cell line identity (STR) | ✓ Confirmed | Matches HEK293T reference (ATCC CRL-3216) |
| Test | Result | Method |
|---|---|---|
| Mycoplasma | ✗ Negative | qPCR or enzymatic assay |
| Sterility — Bacteria & Fungi | ✗ Negative | 14-day culture; no growth |
Certificates of Analysis with lot-specific QC data including flow cytometry data, STR profile, and selection marker confirmation are available upon request. Contact Kulture Scientific with your lot number.
KCL-ENG-302 is shipped as a cryopreserved vial on dry ice. These are adherent HEK293T cells. Follow the steps below upon receipt.
Subculture at 80 to 90% confluence using a cell detachment solution. Split 1:4 to 1:6 every 2 to 3 days. Maintain under appropriate selection antibiotic. Do not maintain cells under selection for more than 4 weeks without re-qualifying ACE2-RFP expression by flow cytometry. Expression may decline after extended culture beyond 20 passages without selection. Full protocols available on the Kulture Scientific protocol page.
KCL-ENG-302 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) Expression in HEK293T Cells | KCL-ENG-302 | 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 — do not refreeze thawed cells; 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
- Expression maintenance — re-qualify ACE2-RFP expression by flow cytometry if cells are maintained under selection for more than 4 weeks, or after extended culture beyond 20 passages
- Certificate of Analysis — lot-specific QC documentation including RFP flow cytometry data, STR profile, selection marker, and sterility results available upon request
For research purposes only. Not for use in diagnostic or therapeutic procedures. Not validated for primary cell models or in vivo injection. 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 — HEK293T cells are derived from human embryonic kidney tissue; all human-derived cell lines require BSL-2 containment under Universal Precautions
- Recombinant DNA — stably integrated ACE2-RFP transgene; standard BSL-2 containment for recombinant human cell lines applies
- ACE2 overexpression — this line highly overexpresses the SARS-CoV-2 entry receptor; when used in pseudovirus or live-virus assays, follow IBC guidance for the viral component
- SV40 Large T antigen — HEK293T cells express the SV40 Large T antigen; requires BSL-2 containment per most institutional biosafety policies
Users must consult their institutional biosafety committee (IBC) prior to use, particularly when used in combination with SARS-CoV-2 pseudovirus or live-virus systems.
- Human-derived material — treat as potentially infectious; work in a certified biosafety cabinet; follow BSL-2 guidelines at all times
- SV40 Large T antigen — present in HEK293T; handle in accordance with institutional BSL-2 requirements for SV40-containing cell lines
- ACE2 expression — overexpresses SARS-CoV-2 entry receptor; when used in pseudovirus 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-302 is a stable HEK293T cell line constitutively expressing full-length human Angiotensin-Converting Enzyme 2 (ACE2) fused with a C-terminal Red Fluorescent Protein (RFP) tag. The ACE2-RFP transgene is stably integrated under a CMV promoter and maintained under Puromycin or G418 (Neomycin) selection — confirm the selection marker for your specific lot with the Certificate of Analysis.
Greater than 90% of cells are RFP-positive by flow cytometry. ACE2 mRNA expression is more than 100-fold over parental HEK293T cells by RT-qPCR. Functional ACE2 enzymatic activity is confirmed by fluorogenic peptide substrate assay (signal-to-background ratio of at least 5:1). Stable single-copy genomic integration is confirmed by PCR or Southern blot. Cell line identity is confirmed by STR profiling against the ATCC CRL-3216 HEK293T reference.
HEK293T is a high-transfection-efficiency derivative of HEK293 widely used in viral pseudotype and protein expression systems. The combination of high ACE2 expression, RFP reporter, and the HEK293T background makes this line well suited for SARS-CoV-2 pseudovirus entry assays, live-cell ACE2 receptor imaging, and drug screening targeting ACE2 activity or Spike/ACE2 interaction. Note: this cell line does not express TMPRSS2; for Spike-primed viral entry, co-express TMPRSS2 or supplement with exogenous trypsin.
DMEM supplemented with 10% heat-inactivated FBS and 1% Pen/Strep. Maintain under Puromycin (1 μg/mL) or G418 (500 μg/mL) selection as appropriate for your lot. Do not maintain cells under selection antibiotic for more than 4 weeks without re-qualifying expression. Subculture at 80 to 90% confluence using a cell detachment solution; split 1:4 to 1:6 every 2 to 3 days. Do not vortex cells.
All lots are quality tested prior to release. The following characterization data is generated for each production lot.
| Parameter | Result | Specification |
|---|---|---|
| ACE2-RFP expression (flow cytometry) | ✓ Positive | ≥90% RFP-positive cells; Ex/Em 558/583 nm |
| ACE2 surface expression | ✓ Positive | High surface expression by immunofluorescence or flow cytometry (anti-ACE2 antibody) |
| ACE2 mRNA expression (RT-qPCR) | ✓ Confirmed | >100-fold over parental HEK293T |
| ACE2 enzymatic activity | ✓ Active | Fluorogenic peptide assay (Mca-APK(Dnp)-OH); signal/background ≥5:1 |
| Genomic integration | ✓ Confirmed | Stable single-copy integration by PCR or Southern blot |
| Post-thaw viability | ✓ Pass | ≥70% by Trypan Blue exclusion |
| Cell line identity (STR) | ✓ Confirmed | Matches HEK293T reference (ATCC CRL-3216) |
| Test | Result | Method |
|---|---|---|
| Mycoplasma | ✗ Negative | qPCR or enzymatic assay |
| Sterility — Bacteria & Fungi | ✗ Negative | 14-day culture; no growth |
Certificates of Analysis with lot-specific QC data including flow cytometry data, STR profile, and selection marker confirmation are available upon request. Contact Kulture Scientific with your lot number.
KCL-ENG-302 is shipped as a cryopreserved vial on dry ice. These are adherent HEK293T cells. Follow the steps below upon receipt.
Subculture at 80 to 90% confluence using a cell detachment solution. Split 1:4 to 1:6 every 2 to 3 days. Maintain under appropriate selection antibiotic. Do not maintain cells under selection for more than 4 weeks without re-qualifying ACE2-RFP expression by flow cytometry. Expression may decline after extended culture beyond 20 passages without selection. Full protocols available on the Kulture Scientific protocol page.
KCL-ENG-302 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) Expression in HEK293T Cells | KCL-ENG-302 | 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 — do not refreeze thawed cells; 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
- Expression maintenance — re-qualify ACE2-RFP expression by flow cytometry if cells are maintained under selection for more than 4 weeks, or after extended culture beyond 20 passages
- Certificate of Analysis — lot-specific QC documentation including RFP flow cytometry data, STR profile, selection marker, and sterility results available upon request
For research purposes only. Not for use in diagnostic or therapeutic procedures. Not validated for primary cell models or in vivo injection. 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 — HEK293T cells are derived from human embryonic kidney tissue; all human-derived cell lines require BSL-2 containment under Universal Precautions
- Recombinant DNA — stably integrated ACE2-RFP transgene; standard BSL-2 containment for recombinant human cell lines applies
- ACE2 overexpression — this line highly overexpresses the SARS-CoV-2 entry receptor; when used in pseudovirus or live-virus assays, follow IBC guidance for the viral component
- SV40 Large T antigen — HEK293T cells express the SV40 Large T antigen; requires BSL-2 containment per most institutional biosafety policies
Users must consult their institutional biosafety committee (IBC) prior to use, particularly when used in combination with SARS-CoV-2 pseudovirus or live-virus systems.
- Human-derived material — treat as potentially infectious; work in a certified biosafety cabinet; follow BSL-2 guidelines at all times
- SV40 Large T antigen — present in HEK293T; handle in accordance with institutional BSL-2 requirements for SV40-containing cell lines
- ACE2 expression — overexpresses SARS-CoV-2 entry receptor; when used in pseudovirus 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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