人ACE2 knockout Caco-2 cell line
Human ACE2 knockout Caco-2 cell line
- Advanced Validation
- 了解详情
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(1 Publication)
ACE2 KO cell line available to order. KO validated by Immunocytochemistry, Western blot. Knockout achieved by using CRISPR/Cas9, Homozygous: 62 bp deletion in exon 2.
查看别名
UNQ868/PRO1885, Angiotensin-converting enzyme homolog, ACE2, ACEH, Metalloprotease MPROT15, Angiotensin-converting enzyme 2, Angiotensin-converting enzyme-related carboxypeptidase, ACE-related carboxypeptidase
- ICC/IF
Lab
Immunocytochemistry/ Immunofluorescence - Human ACE2 knockout Caco-2 cell line (AB273731)
ab272500 staining ACE2 in wild-type Caco2 cells (top panel) and ACE2 knockout Caco2 cells (ab273731) (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab272500 at 10μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
- WB
Lab
Western blot - Human ACE2 knockout Caco-2 cell line (AB273731)
Lanes 1 - 4 : Merged signal (red and green). Green - ab108252 observed at 125 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab108252 was shown to react with ACE2 in Caco-2 wild-type cells in western blot with loss of signal observed in ACE2 knockout cell line ab273731 (knockout cell lysate ab275516). Wild-type and ACE2 knockout Caco-2 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab108252 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4° 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-ACE2 antibody [EPR4435(2)] (<a href='/products/primary-antibodies/ace2-antibody-epr44352-ab108252'>ab108252</a>) at 1/1000 dilution
Lane 1:
Wild-type Caco-2 cell lysate at 30 µg
Lane 2:
ACE2 knockout Caco-2 cell lysate at 30 µg
Lane 3:
Calu-3 cell lysate at 30 µg
Lane 4:
A549 cell lysate at 30 µg
Predicted band size: 104 kDa,60 kDa,76 kDa,92 kDa
Observed band size: 125 kDa,75 kDa
false
- WB
Lab
Western blot - Human ACE2 knockout Caco-2 cell line (AB273731)
Lanes 1 - 4 : Merged signal (red and green). Green - ab108209 observed at 125 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab108209 was shown to react with ACE2 in Caco-2 wild-type cells in western blot with loss of signal observed in ACE2 knockout cell line ab273731 (knockout cell lysate ab275516). Wild-type and ACE2 knockout Caco-2 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab108209 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4° 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-ACE2 antibody [EPR4436] (<a href='/products/primary-antibodies/ace2-antibody-epr4436-ab108209'>ab108209</a>) at 1/1000 dilution
Lane 1:
Wild-type Caco-2 cell lysate at 30 µg
Lane 2:
ACE2 knockout Caco-2 cell lysate at 30 µg
Lane 2:
Western blot - Human ACE2 knockout Caco-2 cell line (ab273731)
Lane 3:
Calu-3 cell lysate at 30 µg
Lane 4:
A549 cell lysate at 30 µg
Predicted band size: 92 kDa
Observed band size: 125 kDa
false
- WB
Lab
Western blot - Human ACE2 knockout Caco-2 cell line (AB273731)
Lanes 1 - 4 : Merged signal (red and green). Green - ab65863 observed at 125 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab65863 was shown to react with ACE2 in Caco-2 wild-type cells in western blot with loss of signal observed in ACE2 knockout cell line ab273731 (knockout cell lysate ab275516). Wild-type and ACE2 knockout Caco-2 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab65863 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4° at 1 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-ACE2 antibody (<a href='/products/primary-antibodies/ace2-antibody-ab65863'>ab65863</a>) at 1 µg/mL
Lane 1:
Wild-type Caco-2 cell lysate at 30 µg
Lane 2:
ACE2 knockout Caco-2 cell lysate at 30 µg
Lane 2:
Western blot - Human ACE2 knockout Caco-2 cell line (ab273731)
Lane 3:
Calu-3 cell lysate at 30 µg
Lane 4:
A549 cell lysate at 30 µg
Predicted band size: 92 kDa
Observed band size: 125 kDa
false
- Sanger seq
Supplier Data
Sanger Sequencing - Human ACE2 knockout Caco-2 cell line (AB273731)
Homozygous : 62 bp deletion in exon 2
搭配使用产品
反应性数据
产品详情
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.
规格
性能和储存信息
基因名称
基因编辑类型
基因编辑方法
敲除验证
合子性
运输条件
推荐的短期储存条件
推荐的长期储存条件
补充信息
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
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.
Pathways
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.
质量控制
STR 分析
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
细胞培养
生物安全等级
EU: 1 US: 1
贴壁/悬浮
Adherent
性别
Male
初始处理指南
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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 70% confluent.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>
传代培养指南
- Do not allow cells to become more than 70% confluent as they will form vacuoles in the cytoplasm.
- Slow to trypsinise, however, prolonged trypsinisation is detrimental to cell viability. Trypsinise until 70% of cells have detached.
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 1x104 - 2x104 cells/cm2 is recommended.
培养基
EMEM + 20% FBS
低温储藏试剂
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
文献 (1)
Recent publications for all applications. Explore the full list and refine your search
Virus evolution 8:veac063 PubMed35919871
2022
Applications
Unspecified application
Species
Unspecified reactive species
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