ATAD2 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 7 bp deletion in exon 1.
AAA nuclear coregulator cancer-associated protein, AAA nuclear coregulator, cancer-associated, ANCCA, ATAD2_HUMAN, ATPase family AAA domain containing 2, ATPase family AAA domain-containing protein 2, CT137, DKFZp667N1320, L16, MGC131938, MGC142216, MGC29843, MGC5254, PRO2000
ATAD2 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 7 bp deletion in exon 1.
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.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
Recommended control: Human wild-type HeLa cell line (ab255448). 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 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.
ATAD2 also known as ATPase family AAA domain-containing protein 2 is a chromatin-associated AAA ATPase. This protein weighs approximately 157 kDa and functions mainly in the cell nucleus. ATAD2 is widely expressed in various tissues with high levels found in the testis indicating its possible role in testicular functions. It shows ATPase activity involving the hydrolysis of ATP which provides energy for chromatin remodeling processes.
ATAD2 plays a critical role in transcription regulation by remodeling chromatin structure. As part of a complex it interacts with histone acetylation marks on chromatin therefore influencing gene expression. The protein is often involved in the regulation of genes relevant to cell cycle progression. Its interaction with transcriptional regulators highlights its importance in controlling proliferation and other cellular processes.
The role of ATAD2 becomes essential in pathways like the cell cycle and the DNA damage response. Through these pathways it interacts with key proteins such as MYC a known regulator of cell growth and it influences the expression of genes involved in cell cycle control. It is important in maintaining genomic stability by ensuring proper chromatin dynamics during DNA replication and repair.
Aberrant expression of ATAD2 associates with cancer progression particularly in breast cancer where it is highly expressed in MCF7 cell lines. It is also implicated in hepatocellular carcinoma where it interacts with cancer-related proteins such as p53 which is a well-known tumor suppressor. These interactions suggest that ATAD2 serves as a potential target for cancer therapies focused on disrupting its interactions or functions in tumor biology.
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Lanes 1- 2: Merged signal (red and green). Green - Anti-ATAD2 antibody [EPR12730] ab176319 observed at 200 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) observed at 50 kDa.
Anti-ATAD2 antibody [EPR12730] ab176319 was shown to react with ATAD2 in wild-type HeLa cells in western blot. The band observed in knockout cell line ab264957 (knockout cell lysate Human ATAD2 knockout HeLa cell lysate ab257359) lane below 200kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and ATAD2 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-ATAD2 antibody [EPR12730] ab176319 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (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-ATAD2 antibody [EPR12730] (Anti-ATAD2 antibody [EPR12730] ab176319) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: ATAD2 CRISPR/Cas9 edited HeLa cell lysate at 20 µg
Lane 2: Western blot - Human ATAD2 knockout HeLa cell line (ab264957)
Performed under reducing conditions.
Predicted band size: 159 kDa
Observed band size: 200 kDa
Homozygous: 7 bp deletion in exon 1.
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