Human ACE2 knockout Caco-2 cell lysate (ab275516)
Overview
-
Product name
Human ACE2 knockout Caco-2 cell lysate
See all ACE2 kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
Caco 2 -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 62 bp deletion in exon 2 -
Passage number
<20 -
Knockout validation
Sanger Sequencing -
Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
-
Notes
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.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.
-
Tested applications
Suitable for: WBmore details
Properties
-
Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab277337 - Human ACE2 knockout Caco 2 cell lysate 1 x 100µg ab277338 - Human wild-type Caco 2 cell lysate 1 x 100µg -
Research areas
-
Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Male
Target
-
Function
Carboxypeptidase which converts angiotensin I to angiotensin 1-9, a peptide of unknown function, and angiotensin II to angiotensin 1-7, a vasodilator. Also able to hydrolyze apelin-13 and dynorphin-13 with high efficiency. May be an important regulator of heart function. In case of human coronaviruses SARS and HCoV-NL63 infections, serve as functional receptor for the spike glycoprotein of both coronaviruses. -
Tissue specificity
Expressed in endothelial cells from small and large arteries, and in arterial smooth muscle cells. Expressed in lung alveolar epithelial cells, enterocytes of the small intestine, Leydig cells and Sertoli cells (at protein level). Expressed in heart, kidney, testis, and gastrointestinal system. -
Sequence similarities
Belongs to the peptidase M2 family. -
Post-translational
modificationsN-glycosylation on Asn-90 may limit SARS infectivity. -
Cellular localization
Secreted and Cell membrane. - Information by UniProt
-
Alternative names
- ACE 2
- ACE related carboxypeptidase
- ACE-related carboxypeptidase
see all
Associated products
-
KO cell lines
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab275516 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
Use at an assay dependent concentration.
|
| Notes |
|---|
|
WB
Use at an assay dependent concentration. |
Images
-
Western blot - Human ACE2 knockout Caco-2 cell lysate (ab275516)
Lane 1: Wild-type Caco-2 cell lysate 30 μg
Lane 2: ACE2 knockout Caco-2 cell lysate 30 μg
Lane 3: Calu-3 cell lysate 30 μg
Lane 4: A549 cell lysate 30 μg
False colour image of Western blot: Anti-ACE2 antibody staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab15348 was shown to bind specifically to ACE2. A band was observed at 120-135 kDa in wild-type Caco-2 cell lysates with no signal observed at this size in ACE2 knockout cell line ab277338 (knockout cell lysate ab275516). To generate this image, wild-type and ACE2 knockout Caco-2 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution. -
Western blot - Human ACE2 knockout Caco 2 cell lysate (ab275516)Lane 1: Wild-type Caco-2 cell lysate 30 ug
Lane 2: ACE2 knockout Caco-2 cell lysate 30 ug
Lane 3: Calu-3 cell lysate 30 ug
Lane 4: A549 cell lysate 30 ug
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 wild-type Caco-2 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°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 (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. -
Western blot - Human ACE2 knockout Caco 2 cell lysate (ab275516)Lane 1: Wild-type Caco-2 cell lysate 30 ug
Lane 2: ACE2 knockout Caco-2 cell lysate 30 ug
Lane 3: Calu-3 cell lysate 30 ug
Lane 4: A549 cell lysate 30 ug
Lanes 1 - 4:Merged signal (red and green). Green - ab108209 observed at 125 kDa. Red - loading controlab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab108209 was shown to react with ACE2 in wild-type Caco-2 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°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 (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. -
Western blot - Human ACE2 knockout Caco 2 cell lysate (ab275516)Lane 1: Wild-type Caco-2 cell lysate 30 ug
Lane 2: ACE2 knockout Caco-2 cell lysate 30 ug
Lane 3: Calu-3 cell lysate 30 ug
Lane 4: A549 cell lysate 30 ug
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 wild-type Caco-2 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°C at 1 ug/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. -
Sanger Sequencing - Human ACE2 knockout Caco 2 cell lysateHomozygous: 62 bp deletion in exon 2
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
-
SDS download
-
Datasheet download
References (0)
ab275516 has not yet been referenced specifically in any publications.