Human ERCC4 knockout HeLa cell line
- Advanced Validation
- What is this?
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Human ERCC4 knockout HeLa cell line available to order. Recommended control: Human wild-type HeLa cell line (ab275466).
View Alternative Names
DNA excision repair protein ERCC-4, DNA repair endonuclease XPF, DNA repair protein complementing XP-F cells, ERCC 11, ERCC 4, Excision repair complementing defective in Chinese hamster, Excision repair cross complementing rodent repair deficiency complementation group 4, FANCQ, XP, group G, XP6, XPF_HUMAN, Xeroderma pigmentosum VI, Xeroderma pigmentosum complementation group F, Xeroderma pigmentosum group F-complementing protein, excision repair cross-complementation group 4
- WB
Lab
Western blot - Human ERCC4 knockout HeLa cell line (AB280068)
False colour image of Western blot : Anti-XPF antibody staining at 1/2000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot ab76948 was shown to bind specifically to XPF. A band was observed at 105 kDa in wild-type HeLa cell lysates with no signal observed at this size in ercc4 knockout cell line ab280068 (knockout cell lysate ab280127). To generate this image wild-type and ercc4 knockout HeLa 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-XPF antibody (<a href='/en-us/products/primary-antibodies/xpf-antibody-ab76948'>ab76948</a>) at 1/2000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
ercc4 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human ERCC4 knockout HeLa cell line (ab280068)
Lane 3:
HEK-293 cell lysate at 20 µg
Lane 4:
Caco-2 cell lysate at 20 µg
Predicted band size: 104 kDa
Observed band size: 105 kDa
false
- Sanger seq
Supplier Data
Sanger Sequencing - Human ERCC4 knockout HeLa cell line (AB280068)
- WB
Supplier Data
Western blot - Human ERCC4 knockout HeLa cell line (AB280068)
False colour image of Western blot : Anti-ercc4 antibody staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, the antibody was shown to bind specifically to ercc4. A band was observed at 105 kDa in wild-type HeLa cell lysates with no signal observed at this size in ercc4 knockout cell line ab280068 (knockout cell lysate ab280127). To generate this image, wild-type and ercc4 knockout HeLa 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°C. 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Human ERCC4 knockout HeLa cell line (ab280068)
false
- WB
Lab
Western blot - Human ERCC4 knockout HeLa cell line (AB280068)
All lanes:
Western blot - Anti-XPF antibody (<a href='/en-us/products/primary-antibodies/xpf-antibody-ab227712'>ab227712</a>) at 1/500 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human ERCC4 knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-ercc4-knockout-hela-cell-lysate-ab280127'>ab280127</a>)
Lane 2:
Western blot - Human ERCC4 knockout HeLa cell line (ab280068)
Lane 2:
ercc4 knockout HeLa cell lysate at 20 µg
Lane 3:
HEK-293 cell lysate at 20 µg
Lane 4:
Caco-2 cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 110 kDa
false
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab275466). 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.
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
The XPF protein collaborates with ERCC1 to form a heterodimeric complex important for nucleotide excision repair (NER). This complex is essential in recognizing and repairing bulky DNA adducts therefore maintaining DNA integrity. The complex performs specific incisions near DNA damage sites removing lesions that frequently occur due to environmental factors like UV radiation and chemical pollutants.
Pathways
XPF functions within the NER and interstrand crosslink repair pathways. In these pathways the XPF-ERCC1 complex coordinates with other proteins like XPA and RPA to accurately excise damaged DNA segments and facilitate repair synthesis. These interactions are important for restoring normal DNA configuration and function after damage thereby preventing mutations and genomic instability.
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