Human MLH1 knockout A549 cell line
- Advanced Validation
- What is this?
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MLH1 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: 70 bp deletion in exon 3.
View Alternative Names
COCA 2, DNA mismatch repair protein Mlh1, FCC 2, HNPCC, HNPCC 2, MGC5172, MLH1_HUMAN, MutL homolog 1, MutL homolog 1 (E. coli), MutL homolog 1 colon cancer nonpolyposis type 2, MutL homolog 1, colon cancer, nonpolyposis type 2 (E. coli), MutL protein homolog 1, MutL, E. coli, homolog of, 1, hMLH 1
- WB
Lab
Western blot - Human MLH1 knockout A549 cell line (AB276105)
False colour image of Western blot : Anti-MLH1 antibody [EPR3894] 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 ab92312 was shown to bind specifically to MLH1. A band was observed at 85 kDa in wild-type A549 cell lysates with no signal observed at this size in MLH1 CRISPR-Cas9 edited cell line ab276105 (CRISPR-Cas9 edited cell lysate ab283566). The band observed in the CRISPR-Cas9 edited lysate lane below 85 kDa is likely to represent a truncated form of MLH1. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image wild-type and MLH1 CRISPR-Cas9 edited A549 cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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-MLH1 antibody [EPR3894] (<a href='/en-us/products/primary-antibodies/mlh1-antibody-epr3894-ab92312'>ab92312</a>) at 1/2000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
MLH1 CRISPR-Cas9 edited A549 cell lysate at 20 µg
Lane 2:
Western blot - Human MLH1 knockout A549 cell line (ab276105)
Lane 2:
Western blot - Human MLH1 knockout A549 cell lysate (<a href='/en-us/products/cell-lysates/human-mlh1-knockout-a549-cell-lysate-ab288239'>ab288239</a>)
Lane 3:
Jurkat cell lysate at 20 µg
Lane 4:
HCT 116 cell lysate at 20 µg
Predicted band size: 85 kDa
Observed band size: 85 kDa
false
- Sanger seq
Lab
Sanger Sequencing - Human MLH1 knockout A549 cell line (AB276105)
70 bp deletion in exon 3
Reactivity data
Product details
Recommended control: Human wild-type A549 cell line (ab255450). 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.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
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
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.
- Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium
F-12K + 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 function of MLH1 involves its role in the mismatch repair (MMR) system. It is part of a complex with PMS2 forming a heterodimer known as MutLα which is essential for the repair process. This complex scans newly synthesized DNA for mispaired bases and initiates repair preserving genomic integrity. The proper function of MLH1 and its interaction with PMS2 ensures that DNA replication errors do not accumulate and cause harmful mutations.
Pathways
MLH1 operates within the mismatch repair pathway and interacts closely with MLH3 and PMS2 proteins. It plays a critical role in the recognition and repair of mismatched bases that occur during DNA replication particularly in the G2 phase of the cell cycle. Through its involvement in the mismatch repair pathway MLH1 is connected to cell cycle regulation and the DNA damage response pathway.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLH1 antibody [EPR3894] (AB92312)](https://content.abcam.com/products/images/mlh1-antibody-epr3894-ab92312--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img423111.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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