ATM KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided.
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- Cancer research communications 3:1731-17422023Discovery and Characterization of ZL-2201, a Potent, Highly Selective, and Orally Bioavailable Small-molecule DNA-PK Inhibitor.Applications:Unspecified applicationReactive species:Unspecified reactive speciesShruti Lal,Neil E Bhola,Bee-Chun Sun,Yuping Chen,Tom Huang,Vivian Morton,Kevin X Chen,Shanghua Xia,Haoyu Zhang,Nehal S Parikh,Qiuping Ye,O Petter Veiby,David I Bellovin,Yuhua JiPubMed 37663435
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
Alternative names
A-T mutated, A-T mutated homolog, AT1, ATC, ATD, ATDC, ATE, ATM serine/threonine kinase, ATM_HUMAN, Ataxia telangiectasia mutated, Ataxia telangiectasia mutated gene, Ataxia telangiectasia mutated homolog, Ataxia telangiectasia mutated homolog (human), DKFZp781A0353, MGC74674, OTTHUMP00000232981, Serine-protein kinase ATM, Serine/threonine-protein kinase ATM, TEL1, TEL1, telomere maintenance 1, homolog, TELO1, Tefu, Telomere fusion protein
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ATM KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided.
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- ATM
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Male
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.
- Do not allow the cell density to exceed 7x104 cells/cm2.
- Culture medium
- F-12K + 10% FBS
- Cryopreservation medium
- Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
ATM also known as Ataxia Telangiectasia Mutated is a protein kinase with a molecular weight of approximately 370 kDa. ATM protein primarily resides in the cell nucleus and functions as a critical regulator of the cell cycle. It plays a significant role in the detection of DNA damage and initiation of repair processes. As part of its mechanical functions ATM phosphorylates serine and threonine residues on various substrates most notably in response to double-strand breaks in DNA. This activity is important for maintaining genomic stability.
- Biological function summary
ATM acts as a coordinator in cellular response to DNA damage highly interacting with multiple components of the DNA repair machinery. It forms a complex with proteins like NBS1 and MRN complex facilitating repair by recruiting and activating other proteins involved in homologous recombination and non-homologous end joining pathways. ATM also modulates p53 activity a primary response factor in cellular stress management linking ATM to control of cell cycle arrest and apoptosis. This positions ATM as an integral part of maintaining cellular integrity in face of genomic insult.
- Pathways
ATM integrates neatly within the DNA damage response and cell cycle control pathways. ATM's operative relationship with the MRN complex and its role in the PI3K-related protein kinase family helps initiate appropriate repair processes upon DNA damage detection. Additionally ATM regulates the activity of proteins such as Chk2 which further propagates signals to p53 influencing decisions between cell cycle arrest and apoptosis. These interactions link ATM closely to essential processes like DNA repair and cell survival highlighting its role in genomic maintenance.
- Associated diseases and disorders
ATM mutations or dysregulation leads to Ataxia Telangiectasia an autosomal recessive disorder characterized by neurodegeneration immune deficiencies and cancer predisposition. ATM dysfunction also connects to cancer development particularly breast cancer where it transmits signals involving BRCA1 contributing to DNA repair through homologous recombination. Understanding ATM dynamics and related pathways has important implications for developing therapeutic strategies to manage or mitigate effects associated with its dysfunction.
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4 product images
Western blot - Human ATM knockout A549 cell line (ab276095)
False colour image of Western blot: Anti-ATM antibody [EPR20100] - ChIP Grade staining at 1/1000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-ATM antibody [EPR20100] - ChIP Grade ab201022 was shown to bind specifically to ATM. A band was observed at 350 kDa in wild-type A549 cell lysates with no signal observed at this size in ATM knockout cell line ab276095 (knockout cell lysate ab283834). To generate this image wild-type and ATM knockout A549 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°. 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 (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) at 1/20000 dilution.
All lanes: Western blot - Anti-ATM antibody [EPR20100] - ChIP Grade (Anti-ATM antibody [EPR20100] - ChIP Grade ab201022) at 1/1000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: ATM knockout A549 cell lysate at 20 µg
Lane 2: Western blot - Human ATM knockout A549 cell line (ab276095)
Lane 3: HEK-293 cell lysate at 20 µg
Lane 4: U-2 OS cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 351 kDa
Observed band size: 350 kDa
Western blot - Human ATM knockout A549 cell line (ab276095)
False colour image of Western blot: Anti-ATM antibody [EPR17059] staining at 1/2000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-ATM antibody [EPR17059] ab199726 was shown to bind specifically to ATM. A band was observed at 350 kDa in wild-type A549 cell lysates with no signal observed at this size in ATM knockout cell line ab276095 (knockout cell lysate ab283834). To generate this image wild-type and ATM knockout A549 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°. 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 (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) at 1/20000 dilution.
All lanes: Western blot - Anti-ATM antibody [EPR17059] (Anti-ATM antibody [EPR17059] ab199726) at 1/2000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: ATM knockout A549 cell lysate at 20 µg
Lane 2: Western blot - Human ATM knockout A549 cell line (ab276095)
Lane 3: HEK-293 cell lysate at 20 µg
Lane 4: U-2 OS cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 351 kDa
Observed band size: 350 kDa
Western blot - Human ATM knockout A549 cell line (ab276095)
False colour image of Western blot: Anti-ATM antibody [Y170] staining at 1/1000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-ATM antibody [Y170] ab32420 was shown to bind specifically to ATM. A band was observed at 350 kDa in wild-type A549 cell lysates with no signal observed at this size in ATM knockout cell line ab276095 (knockout cell lysate ab283834). To generate this image wild-type and ATM knockout A549 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°. 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 (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) at 1/20000 dilution.
All lanes: Western blot - Anti-ATM antibody [Y170] (Anti-ATM antibody [Y170] ab32420) at 1/1000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: ATM knockout A549 cell lysate at 20 µg
Lane 2: Western blot - Human ATM knockout A549 cell line (ab276095)
Lane 3: HEK-293 cell lysate at 20 µg
Lane 4: U-2 OS cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 351 kDa
Observed band size: 350 kDa
Sanger Sequencing - Human ATM knockout A549 cell line (ab276095)
Knockout achieved by using CRISPR/Cas9, Homozygous: 28 bp deletion in exon 3
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