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HEK293/ACE2 Stable Cell Line

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).

HEK293/ACE2 Stable Cell Line

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.

HEK293/ACE2 Stable Cell Line

FACS analysis histogram shows ACE2 expression in HEK293/ACE2 stable cell line (Cat. No. M00770) compared with negative control. HEK293/ACE2 cells are plated at 3E5+ cells /well in 100ul PBS in 96-well plate, incubated with 10 µg/ml S-RBD-His (GenScript, Cat. No. Z03479) on ice for 1 hour, or left untreated. Both groups are then washed with PBS, incubated with Anti-His 2nd antibody (GenScript, Cat. No. A01800) at 1:200 on ice for 1 hour, washed with PBS, and then analyzed on flow cytometer. 

HEK293/ACE2 Stable Cell Line

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.

HEK293/ACE2 Stable Cell Line

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

HEK293/ACE2 Stable Cell Line

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.
M00770
$8,500.00

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Overview
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. 

Product Introduction
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 Conditions
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)

Background
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.

Examples
  • HEK293/ACE2 Stable Cell Line
  • HEK293/ACE2 Stable Cell Line

    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).

  • HEK293/ACE2 Stable Cell Line
  • HEK293/ACE2 Stable Cell Line

    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.

  • HEK293/ACE2 Stable Cell Line
  • HEK293/ACE2 Stable Cell Line

    FACS analysis histogram shows ACE2 expression in HEK293/ACE2 stable cell line (Cat. No. M00770) compared with negative control. HEK293/ACE2 cells are plated at 3E5+ cells /well in 100ul PBS in 96-well plate, incubated with 10 µg/ml S-RBD-His (GenScript, Cat. No. Z03479) on ice for 1 hour, or left untreated. Both groups are then washed with PBS, incubated with Anti-His 2nd antibody (GenScript, Cat. No. A01800) at 1:200 on ice for 1 hour, washed with PBS, and then analyzed on flow cytometer. 

  • HEK293/ACE2 Stable Cell Line
  • HEK293/ACE2 Stable Cell Line

    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.

  • HEK293/ACE2 Stable Cell Line
  • HEK293/ACE2 Stable Cell Line

    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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