ACE2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 71 bp deletion in exon 2.
ACE-related carboxypeptidase, ACE2_HUMAN, ACEH, Angiotensin I Converting Enzyme (peptidyl dipeptidase A) 2, Angiotensin I converting enzyme 2, Angiotensin converting enzyme 2, Angiotensin converting enzyme like protein, Angiotensin-converting enzyme homolog, DKFZP434A014, EC 3.4.17, OTTHUMP00000022963, Processed angiotensin-converting enzyme 2, metalloprotease MPROT 15
ACE2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 71 bp deletion in exon 2.
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
The ACE2 protein also known as angiotensin-converting enzyme 2 is an essential component in the renin-angiotensin system. It functions mechanically by converting the hormone angiotensin II to angiotensin-(1-7) which helps regulate blood pressure and fluid balance. The molecular weight of ACE2 is approximately 120 kDa. This protein is expressed in various tissues particularly the lungs heart kidneys and gastrointestinal tract. In cultured cells like Caco-2 cells researchers often study this expression.
The ACE2 protein plays an important role in the regulation of cardiovascular and renal functions. It is a single-pass type I membrane protein and its activity reduces inflammation and oxidative stress in cells. ACE2 does not function as part of a larger protein complex but its enzymatic conversion has a substantial impact on reducing the effects of angiotensin II in the body leading to vasodilation and decreased blood pressure.
ACE2 involvement is significant in the renin-angiotensin system and the kallikrein-kinin system. These pathways are essential for maintaining cardiovascular homeostasis. In the renin-angiotensin system ACE2 works in opposition to angiotensin-converting enzyme (ACE) balancing the effects through the production of angiotensin-(1-7) from angiotensin II. Additionally ACE2 interacts indirectly with proteins like angiotensin receptor type 1 (AT1) and angiotensin receptor type 2 (AT2) ensuring proper signaling and physiological responses.
ACE2 links closely with conditions such as hypertension and COVID-19. Increased activity of angiotensin II due to low ACE2 levels contributes to hypertension. In infectious disease SARS-CoV-2 virus responsible for COVID-19 uses ACE2 as an entry receptor to initiate infection in host cells. This interaction highlights the importance of ACE2 in disease pathogenesis and has prompted interest in ACE2 as a potential therapeutic target.
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Lanes 1 - 4: Merged signal (red and green). Green - Anti-ACE2 antibody [EPR4436] ab108209 observed at 130 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
Anti-ACE2 antibody [EPR4436] ab108209 was shown to react with ACE2 in wild-type HepG2 cells in western blot with loss of signal observed in ACE2 knockout cell line ab273733 (knockout cell lysate Human ACE2 knockout Hep G2 cell lysate ab275495). Wild-type and ACE2 knockout HepG2 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with Anti-ACE2 antibody [EPR4436] ab108209 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-ACE2 antibody [EPR4436] (Anti-ACE2 antibody [EPR4436] ab108209) at 1/1000 dilution
Lane 1: Wild-type HepG2 cell lysate at 30 µg
Lane 2: ACE2 knockout HepG2 cell lysate at 30 µg
Lane 2: Western blot - Human ACE2 knockout Hep G2 cell line (ab273733)
Lane 3: Calu-3 cell lysate at 30 µg
Lane 4: A549 cell lysate at 30 µg
Performed under reducing conditions.
Predicted band size: 92 kDa
Observed band size: 130 kDa
Lanes 1 - 4: Merged signal (red and green). Green - Anti-ACE2 antibody ab65863 observed at 130 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
Anti-ACE2 antibody ab65863 was shown to react with ACE2 in wild-type HepG2 cells in western blot with loss of signal observed in ACE2 knockout cell line ab273733 (knockout cell lysate Human ACE2 knockout Hep G2 cell lysate ab275495). Wild-type and ACE2 knockout HepG2 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with Anti-ACE2 antibody ab65863 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-ACE2 antibody (Anti-ACE2 antibody ab65863) at 1 µg/mL
Lane 1: Wild-type HepG2 cell lysate at 30 µg
Lane 2: ACE2 knockout HepG2 cell lysate at 30 µg
Lane 2: Western blot - Human ACE2 knockout Hep G2 cell line (ab273733)
Lane 3: Calu-3 cell lysate at 30 µg
Lane 4: A549 cell lysate at 30 µg
Performed under reducing conditions.
Predicted band size: 92 kDa
Observed band size: 130 kDa
Allele 1: 71 bp deletion in exon 2.
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