| Product Description |
Recombinant HEK293 cells
stably express Angiotensin-converting enzyme 2 (ACE2) on the cell surface. The
surface expression of ACE2 is validated by FACS analysis. This stable cell line
product is designed for cell-based binding assays that measure the binding
affinity and stability of antibody based biologics binding with ACE2, and can
be utilized for SARS-Cov-2 pseudovirus neutralization assay for screening
neutralizing antibodies.
|
| Applications |
Cell-based binding assay; neutralization assay; use as
immunogen for antibody development.
|
| Expressed Gene |
Codon Optimized from NM_021804.2; no expressed tags
|
| Target Protein |
NP_068576.1
|
| Host Cell |
HEK293
|
| Quantity |
2 vials of frozen cells (>1X106 per
vial in 1 ml)
|
| Storage |
Store cells in liquid nitrogen immediately upon
receipt. Thaw and recover cells within one year from the date received.
|
| Mycoplasma Status |
Negative.
The mycoplasma test was performed with MycoAlert? PLUS Mycoplasma Detection Kit
(Cat. No. LT07-318, Lonza).
|
| Culture Properties |
Adherent
|
| Freeze Medium |
95% complete growth medium, 5% DMSO (Cat. No. D2650,
Sigma)
|
| Complete Growth Medium |
DMEM (Cat. No. 10566-016, Gibco), 10% FBS (Cat. No. 10099-141,
Gibco)
|
| Culture Medium |
DMEM, 10% FBS, 250 μg/ml Hygromycin B (Cat. No. 10687010,
Invitrogen)
|
| Gene Synonyms |
Angiotensin-converting enzyme 2, ACEH
|
| Background |
Angiotensin-converting
enzyme 2 (ACE2), discovered as a homologue of ACE, acts as its physiological
counterbalance providing homeostatic regulation of circulating angiotensin II
(Ang II) levels. ACE2 is a zinc metalloenzyme and carboxypeptidase located as
an ectoenzyme on the surface of endothelial and other cells. While its primary
substrate appears to be Ang II, it can hydrolyze a number of other
physiological substrates. Additionally, ACE2 functions in other noncatalytic
cellular roles including the regulation of intestinal neutral amino acid
transport. It also serendipitously acts as the receptor for the severe acute
respiratory syndrome virus. Upregulation of ACE2 expression and function is
increasingly recognized as a potential therapeutic strategy in hypertension and
cardiovascular disease, diabetes, lung injury, and fibrotic disorders. ACE2 is
regulated at multiple levels including transcriptional, posttranscriptional
(miRNA and epigenetic), and posttranslational through its shedding from the
cell surface.
|
Figure 1: FACS analysis of cell surface expression of ACE2 on HEK293 cells.
HEK293/ACE2 cells (red) and negative control HEK293 cells (green) were stained with FITC conjugated SARS-CoV-2 Spike protein (RBD, His Tag) (Cat. No. Z03479, GenScript).
Figure 2: Cell-based binding assay of SARS-CoV-2 spike protein RBD with HEK293/ACE2 cells.
HEK293/ACE2 cells were incubated with SARS-CoV-2 Spike protein (RBD, His Tag) (Cat. No. Z03479, GenScript) in serial dilution. Then the cells were stained with THE? His Tag Antibody [iFluor 488], mAb, Mouse (Cat. No. A01800, GenScript) and analyzed with flow cytometer. FACS analysis shows the EC50 of spike protein RBD binding with ACE2 on HEK293 cells is 1.071 ug/ml.
Figure 3: Neutralization assay of SARS-CoV-2 spike protein RBD by ACE2-Fc fusion proteins on HEK293/ACE2 cells.
HEK293/ACE2 cells were incubated with 1 μg/ml SARS-CoV-2 Spike protein (RBD, His Tag) (Cat. No. Z03479, GenScript), and ACE2-Fc fusion protein (Cat. No. Z03484, GenScript) in serial dilution. Then the cells were stained with THE? His Tag Antibody [iFluor 488], mAb, Mouse (Cat. No. A01800, GenScript) and analyzed with flow cytometer.
Figure 5: FACS analysis of cell surface expression of ACE2 on HEK293 cells from different passages.
The HEK293/ACE2 cells and the negative control HEK293 cells were stained with FITC conjugated SARS-CoV-2 Spike protein (RBD, His Tag) (Cat. No. Z03479, GenScript). P8, P13, P17 and P21 represent cell passage numbers.
Figure 4: SARS-CoV-2 (B.1.1.529, Omicron) pseudovirus transfection on HEK293/ACE2 cells.
SARS-CoV-2 pseudovirus with S glycoprotein as the envelope protein. The pseudovirus entry was initiated by recognition and binding between RBD and ACE2 on HEK293 cells
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