Human TRIM29 knockout HeLa cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human TRIM29 knockout HeLa cell line (AB265878)
Lanes 1-4 : Merged signal (red and green). Green - ab108627 observed at 74 kDa. Red - loading control ab8245 observed at 37 kDa.
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 ab257770) was used. Wild-type and TRIM29 knockout samples were subjected to SDS-PAGE. ab108627 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-TRIM29 antibody [EPR3494] (<a href='/en-us/products/primary-antibodies/trim29-antibody-epr3494-ab108627'>ab108627</a>) 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 2:
Western blot - Human TRIM29 knockout HeLa cell line (ab265878)
Lane 3:
A431 cell lysate at 20 µg
Lane 4:
Daudi 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 1/20000 dilution
Predicted band size: 65 kDa
Observed band size: 74 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human TRIM29 knockout HeLa cell line (AB265878)
Homozygous : Insertion of the selection cassette in exon 1.
Complementary Products
Primary Antibodies
AB247707
Anti-TRIM29 antibody [EPR3494] - BSA and Azide free
RabMAb|Recombinant|KO Validated
![Western blot - Anti-TRIM29 antibody [EPR3494] - BSA and Azide free (AB247707)](https://content.abcam.com/products/images/trim29-antibody-epr3494-bsa-and-azide-free-ab247707--western-blot-img2156.jpg)
- 0
- 0
Reactivity data
Product details
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
Zygosity
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
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.
Pathways
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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