JavaScript is disabled in your browser. Please enable JavaScript to view this website.
AB273731

Human ACE2 knockout Caco-2 cell line

Be the first to review this product! Submit a review

|

(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.
5 Images
Immunocytochemistry/ Immunofluorescence - Human ACE2 knockout Caco-2 cell line (AB273731)
  • 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).

Western blot - Human ACE2 knockout Caco-2 cell line (AB273731)
  • 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='/en-us/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

Western blot - Human ACE2 knockout Caco-2 cell line (AB273731)
  • 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='/en-us/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

Western blot - Human ACE2 knockout Caco-2 cell line (AB273731)
  • 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='/en-us/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 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: 104 kDa,60 kDa,76 kDa,92 kDa

Observed band size: 125 kDa,75 kDa

false

Sanger Sequencing - Human ACE2 knockout Caco-2 cell line (AB273731)
  • Sanger seq

Supplier Data

Sanger Sequencing - Human ACE2 knockout Caco-2 cell line (AB273731)

Homozygous : 62 bp deletion in exon 2

Key facts

Cell type

Caco-2

Species or organism

Human

Tissue

Colon

Form

Liquid

form

Knockout validation

Immunocytochemistry,Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 62 bp deletion in exon 2

Disease

Adenocarcinoma

style
left-column

Complementary Products

Primary Antibodies

AB108252

Anti-ACE2 antibody [EPR4435(2)]

BOND RX™ Validated|RabMAb|Recombinant|KO Validated|20ul selling size

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody [EPR4435(2)] (AB108252)

  • 4
  • 10
Secondary Antibodies

AB150077

Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)

Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (AB150077)

  • 5
  • 20
Proteins & Peptides

AB151852

Recombinant Human ACE2 protein

Western blot - Recombinant Human ACE2 protein (AB151852)

  • 0
  • 0
Primary Antibodies

AB109131

Anti-TMPRSS2 antibody [EPR3862]

RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMPRSS2 antibody [EPR3862] (AB109131)

  • 5
  • 3
style
left-column

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
style
left-column

Product details

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.

style
left-column

What's included?

{ "values": { "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab273731-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab273731 Human ACE2 knockout Caco-2 cell line", "number":"AB273731-CMP01", "productcode":"" } ] } } }
style
left-column

Properties and storage information

Gene name
ACE2
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Immunocytochemistry, Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
style
left-column

Handling procedures

Initial handling guidelines

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 70% confluent.
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.

Subculture guidelines
  • 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.
Culture medium

EMEM + 20% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

style
left-column

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.
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.

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.
style
left-column
style
left-column

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

style
left-column

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

style
left-column

Product protocols

style
left-column

Target data

See full target information ACE2
style
left-column

Publications (1)

Recent publications for all applications. Explore the full list and refine your search

Virus evolution 8:veac063 PubMed35919871

2022

Evolution of ACE2-independent SARS-CoV-2 infection and mouse adaption after passage in cells expressing human and mouse ACE2.

Applications

Unspecified application

Species

Unspecified reactive species

Kexin Yan,Troy Dumenil,Bing Tang,Thuy T Le,Cameron R Bishop,Andreas Suhrbier,Daniel J Rawle
View all publications
style
left-column
style
left-column
style
right-column

Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com