Human ADAR (ADAR1) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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Human ADAR (ADAR1) knockout HEK-293T cell line available to order. Recommended control: Human wild-type HEK293T cell line (ab255449).
View Alternative Names
136 kDa double-stranded RNA-binding protein, ADAR, AGS6, AV242451, Adenosine deaminase RNA specific, Adenosine deaminase RNA specific 1, Adenosine deaminase acting on RNA 1 A, Adenosine deaminase that act on RNA, DRADA, DSRAD_HUMAN, Double-stranded RNA-specific adenosine deaminase, Double-stranded RNA-specific editase Adar, Dsh, EC 3.5.4.-, G1P1, Ifi4 protein, Interferon induced protein 4, Interferon-inducible protein 4, K88DSRBP, P136, Pre-mRNA adenosine deaminase, RNA adenosine deaminase 1, RNA-editing deaminase 1, RNA-editing enzyme 1, dsRNA adenosine deaminase, mZaADAR
- ICC/IF
Supplier Data
Immunocytochemistry/ Immunofluorescence - Human ADAR (ADAR1) knockout HEK-293T cell line (AB266846)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized ADAR KO HEK293T (ab266846) cells labelling ADAR1 with ab307585 at 1/500 (1.08 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing increased nuclear and cytoplasmic staining in parental HEK293T cells treated with IFN alpha 1 (human) (10 ng/ml) for 16 hours, and no staining in treated ADAR KO HEK293T cells. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.
- WB
Lab
Western blot - Human ADAR (ADAR1) knockout HEK-293T cell line (AB266846)
Lanes 1-3 : Merged signal (red and green). Green - ab126745 observed at 130 kDa. Red - loading control ab8245 observed at 36 kDa.
ab126745 Anti-ADAR1 antibody [EPR7033] was shown to specifically react with ADAR1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266846 (knockout cell lysate ab257131) was used. Wild-type and ADAR1 knockout samples were subjected to SDS-PAGE. ab126745 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and 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-ADAR1 antibody [EPR7033] (<a href='/en-us/products/primary-antibodies/adar1-antibody-epr7033-ab126745'>ab126745</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
ADAR knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human ADAR (ADAR1) knockout HEK-293T cell line (ab266846)
Lane 3:
HeLa cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 136 kDa,19 kDa,29 kDa,35 kDa,36 kDa
Observed band size: 130 kDa,20 kDa,29 kDa,35 kDa,36 kDa
false
- WB
Supplier Data
Western blot - Human ADAR (ADAR1) knockout HEK-293T cell line (AB266846)
Blocking and diluting buffer and concentration : 5% NFDM/TBST Performed under reducing conditions. In Western blot, ab307585 was shown to bind specifically to ADAR1. A band was observed at 150 kDa in wild-type HEK-293T cell lysates whereas no signal observed at this size in ADAR1 knockout cell line ab266846 (knockout cell lysate ab257131). Exposure time : 59 seconds
All lanes:
Western blot - Anti-ADAR1 antibody [EPR25431-60] (<a href='/en-us/products/primary-antibodies/adar1-antibody-epr25431-60-ab307585'>ab307585</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg
Lane 2:
ADAR1 knockout HEK-293T whole cell lysate at 20 µg
Lane 2:
Western blot - Human ADAR (ADAR1) knockout HEK-293T cell line (ab266846)
Lane 2:
Western blot - Human ADAR (ADAR1) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-adar-adar1-knockout-hek-293t-cell-lysate-ab257131'>ab257131</a>)
Lane 3:
HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 4:
Ramos (human burkitts lymphoma b lymphocyte) whole cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution
Observed band size: 150 kDa
false
Exposure time: 59s
- Sanger seq
Unknown
Sanger Sequencing - Human ADAR (ADAR1) knockout HEK-293T cell line (AB266846)
Allele-2 : 1 bp insertion in exon 2.
- Sanger seq
Lab
Sanger Sequencing - Human ADAR (ADAR1) knockout HEK-293T cell line (AB266846)
Sequencing chromatogram displaying sequence edit in exon 2
- Sanger seq
Unknown
Sanger Sequencing - Human ADAR (ADAR1) knockout HEK-293T cell line (AB266846)
Allele-1 : 17 bp deletion in exon2
Reactivity data
Product details
Recommended control: Human wild-type HEK293T cell line (ab255449). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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 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.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
ADAR1 plays a role in the regulation of RNA molecules affecting their stability and translation. It is linked to the editosome complex working alongside other proteins to perform RNA editing tasks. By modifying the coding potential of mRNAs ADAR1 contributes to the diversity of proteomes and helps manage the responses to viral RNAs giving the immune system tools to recognize endogenous and exogenous RNA.
Pathways
ADAR1 is significant in the interferon signaling pathway and RNA processing pathways. It operates in coordination with proteins like PKR which is involved in the response to viral infections. ADAR1 ensures that the immune response is not directed against the self highlighting its role in the regulation of the innate immune system. These pathways are critical in maintaining homeostasis and preventing unchecked immune responses.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Product protocols
- Visit the General protocols
- Visit the Troubleshooting
Target data
Product promise
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