Human POLL (DNA Polymerase lambda) knockout HeLa cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human POLL (DNA Polymerase lambda) knockout HeLa cell line (AB264754)
Lanes 1-3 : Merged signal (red and green). Green - ab172974 observed at 75 kDa. Red - loading control ab8245 observed at 37 kDa.
ab172974 Anti-DNA Polymerase lambda/Polk antibody [EPR7519(2)] was shown to specifically react with DNA Polymerase lambda/Polk in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264754 (knockout cell lysate ab258595) was used. Wild-type and DNA Polymerase lambda/Polk knockout samples were subjected to SDS-PAGE. ab172974 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 10000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-DNA Polymerase lambda antibody [EPR7519(2)] (<a href='/en-us/products/primary-antibodies/dna-polymerase-lambda-antibody-epr75192-ab172974'>ab172974</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
POLL knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human POLL (DNA Polymerase lambda) knockout HeLa cell line (ab264754)
Lane 3:
A549 cell lysate at 20 µg
Predicted band size: 63 kDa
Observed band size: 75 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human POLL (DNA Polymerase lambda) knockout HeLa cell line (AB264754)
Allele-3 : Insertion of the selection cassette in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human POLL (DNA Polymerase lambda) knockout HeLa cell line (AB264754)
Allele-2 : Insertion of the selection cassette in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human POLL (DNA Polymerase lambda) knockout HeLa cell line (AB264754)
Allele-1 : 1 bp deletion in exon 2.
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Primary Antibodies
AB172974
Anti-DNA Polymerase lambda antibody [EPR7519(2)]
RabMAb|Recombinant|KO Validated
![Western blot - Anti-DNA Polymerase lambda antibody [EPR7519(2)] (AB172974)](https://content.abcam.com/products/images/dna-polymerase-lambda-antibody-epr75192-ab172974--western-blot-img105148.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
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
DNA polymerase lambda acts as a gap-filling enzyme during DNA repair processes especially in base excision repair and non-homologous end joining. This polymerase does not typically function within a larger complex but requires accessory factors to perform its repair functions. It maintains genomic stability by providing an accurate extension of DNA strands during repair ensuring the correct rejoining of DNA. Furthermore it also exhibits a lyase activity that can remove 5'-deoxyribose phosphate residues.
Pathways
DNA polymerase lambda is actively involved in the base excision repair and non-homologous end joining pathways. These pathways are important for fixing DNA single-strand breaks and double-strand breaks respectively. Pol λ works alongside proteins like XRCC1 in the short-patch base excision repair pathway and interacts with KU70/80 in non-homologous end joining reinforcing its multifaceted role in maintaining DNA integrity under varying genotoxic stress conditions.
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'.
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