TRIM29 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1.
HeLa
Human
Cervix
Liquid
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1
ATDC, Ataxia telangiectasia group D-associated protein, FLJ36085, TRI29_HUMAN, Tripartite motif containing 29, Tripartite motif protein 29, Tripartite motif protein TRIM29, Tripartite motif-containing protein 29
TRIM29 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1.
HeLa
Human
Cervix
Liquid
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1
Adenocarcinoma
TRIM29
Knockout
CRISPR technology
Sanger Sequencing, Western blot
Homozygous
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
EU: 2 US: 2
Adherent
Female
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.
DMEM (High Glucose) + 10% FBS
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Dry Ice
-196°C
Recommended control: Human wild-type HeLa cell line (Human wild-type HeLa cell line ab255928). 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.
This supplementary information is collated from multiple sources and compiled automatically.
The tripartite motif-containing protein 29 (TRIM29) also known as ATDC is a protein with a molecular mass of approximately 66 kDa. TRIM29 is part of the TRIM family characterized by its RING finger motif B-box type 1 and 2 domains and a coiled-coil region which are significant for its involvement in ubiquitination processes. Researchers have found TRIM29 expression largely in epithelial tissues exhibiting varied levels of expression depending on tissue type.
TRIM29 functions as an essential regulator of DNA damage responses and cellular proliferation. It is not found as part of any larger protein complex but interacts with various other proteins to carry out its function. TRIM29 can influence the activity of key molecules involved in DNA repair particularly through its interaction with DNA-dependent protein kinase catalytic subunit (DNA-PKcs) which enhances cell survival after genotoxic stress. It participates in ensuring proper functioning of cell cycle checkpoints and modulation of apoptosis.
TRIM29 participates in the DNA damage response pathway and the WNT signaling pathway. It interacts with proteins such as β-catenin which influences the transcriptional regulation of target genes involved in cell proliferation. Through these interactions TRIM29 influences cellular responses to DNA damage and the regulation of cell growth linking it to critical pathways that maintain genomic stability and cell integrity.
Aberrant TRIM29 expression correlates with various cancers including breast and colorectal cancers. In breast cancer TRIM29 affects the pathways involving p53 a known tumor suppressor protein. In colorectal cancer its overexpression is associated with enhanced cancer cell proliferation and survival often in conjunction with KRAS mutations. TRIM29's regulatory role in these pathways highlights its potential as a biomarker and a therapeutic target in oncology.
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Lanes 1-4: Merged signal (red and green). Green - Anti-TRIM29 antibody [EPR3494] ab108627 observed at 74 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
Anti-TRIM29 antibody [EPR3494] ab108627 Anti-TRIM29 antibody [EPR3494] was shown to specifically react with TRIM29 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265878 (knockout cell lysate Human TRIM29 knockout HeLa cell lysate ab257770) was used. Wild-type and TRIM29 knockout samples were subjected to SDS-PAGE. Anti-TRIM29 antibody [EPR3494] ab108627 and Anti-GAPDH antibody [6C5] - Loading Control (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 (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-TRIM29 antibody [EPR3494] (Anti-TRIM29 antibody [EPR3494] ab108627) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: TRIM29 knockout HeLa cell lysate at 20 µg
Lane 3: A431 cell lysate at 20 µg
Lane 4: Daudi cell lysate at 20 µg
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 65 kDa
Observed band size: 74 kDa
Lanes 1-4: Merged signal (red and green). Green - Anti-TRIM29 antibody [EPR3494] ab108627 observed at 74 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
Anti-TRIM29 antibody [EPR3494] ab108627 Anti-TRIM29 antibody [EPR3494] was shown to specifically react with TRIM29 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265878 (knockout cell lysate Human TRIM29 knockout HeLa cell lysate ab257770) was used. Wild-type and TRIM29 knockout samples were subjected to SDS-PAGE. Anti-TRIM29 antibody [EPR3494] ab108627 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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.
Homozygous: Insertion of the selection cassette in exon 1.
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