POLL KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and Insertion of the selection cassette in exon 2.
BETA N, DNA directed DNA polymerase lambda, DNA polymerase beta-2, DNA polymerase kappa, DNA polymerase kappa DNA polymerase beta N, DNA polymerase lambda, DNA polymerase lamda2, DPOLL_HUMAN, EC 2.7.7.7,EC 4.2.99., FLJ46002, OTTHUMP00000020321, OTTHUMP00000020323, OTTHUMP00000059179, POL KAPPA, POLL, Pol Lambda, Pol beta2, Polymerase DNA directed lambda
POLL KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and Insertion of the selection cassette in exon 2.
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.
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.
DNA polymerase lambda also known as pol λ or DNA pol λ is a DNA polymerase protein involved in DNA repair and replication processes. It has a molecular weight of approximately 65 kDa. DNA polymerase lambda is expressed in various tissues but shows a high concentration in testis and thymus indicating its role in germ cells and immune response. The enzyme exhibits both DNA polymerase and terminal transferase activities facilitating DNA synthesis across gaps and lesions.
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.
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.
DNA polymerase lambda is linked to conditions such as cancer and immunodeficiencies where its dysfunction can lead to genomic instability. The loss or aberrant function of DNA pol λ may result in ineffective DNA repair contributing to cancer development due to accumulated DNA damage. It can also interact with tumor suppressor p53 highlighting its involvement in pathways that prevent oncogenesis. Additionally impairments in its function can contribute to immunodeficiency disorders as proper DNA repair is essential for the development and function of the immune system.
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Anti-DNA Polymerase lambda antibody [EPR7519(2)] 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 Human POLL (DNA Polymerase lambda) knockout HeLa cell lysate ab258595) was used. Wild-type and DNA Polymerase lambda/Polk knockout samples were subjected to SDS-PAGE. Anti-DNA Polymerase lambda antibody [EPR7519(2)] ab172974 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 10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-DNA Polymerase lambda antibody [EPR7519(2)] (Anti-DNA Polymerase lambda antibody [EPR7519(2)] ab172974) 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
Performed under reducing conditions.
Predicted band size: 63 kDa
Observed band size: 75 kDa
Allele-3: Insertion of the selection cassette in exon 2.
Allele-1: 1 bp deletion in exon 2.
Allele-2: Insertion of the selection cassette in exon 2.
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