Human MLH1 knockout HeLa cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Supplier Data
Western blot - Human MLH1 knockout HeLa cell line (AB267223)
Lanes 1-4 : Merged signal (red and green). Green - ab108622 observed at 90 kDa. Red - loading control ab8245 observed at 37 kDa.
ab108622 Anti-MLH1 antibody [EPR3893] was shown to specifically react with MLH1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab267223 (knockout cell lysate ab257172) was used. Wild-type and MLH1 knockout samples were subjected to SDS-PAGE. ab108622 and Anti-GAPDH antibody [6C5] - Loading Control were incubated overnight at 4° at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-MLH1 antibody [EPR3893] (<a href='/en-us/products/primary-antibodies/mlh1-antibody-epr3893-ab108622'>ab108622</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
MLH1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human MLH1 knockout HeLa cell line (ab267223)
Lane 3:
Jurkat cell lysate at 20 µg
Lane 4:
HCT116 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: 85 kDa
Observed band size: 85 kDa
false
- WB
Supplier Data
Western blot - Human MLH1 knockout HeLa cell line (AB267223)
Lanes 1-4 : Merged signal (red and green). Green - ab92312 observed at 90 kDa. Red - loading control ab8245 observed at 37 kDa.
ab92312 Anti-MLH1 antibody [EPR3894] was shown to specifically react with MLH1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab267223 (knockout cell lysate ab257172) was used. Wild-type and MLH1 knockout samples were subjected to SDS-PAGE. ab92312 and Anti-GAPDH antibody [6C5] - Loading Control were incubated overnight at 4° at 1 in 10000 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-MLH1 antibody [EPR3894] (<a href='/en-us/products/primary-antibodies/mlh1-antibody-epr3894-ab92312'>ab92312</a>) at 1/10000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
MLH1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human MLH1 knockout HeLa cell line (ab267223)
Lane 3:
Jurkat cell lysate at 20 µg
Lane 4:
HCT116 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: 85 kDa
Observed band size: 85 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human MLH1 knockout HeLa cell line (AB267223)
Allele-2 : 4 bp deletion in exon 12.
- Sanger seq
Unknown
Sanger Sequencing - Human MLH1 knockout HeLa cell line (AB267223)
Allele-1 : 10 bp deletion in exon12
- Cell Culture
Unknown
Cell Culture - Human MLH1 knockout HeLa cell line (AB267223)
Representative images of MLH1 knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human MLH1 knockout HeLa cell line (AB267223)
Allele-3 : 1 bp deletion in exon 12.
Complementary Products
Primary Antibodies
AB92312
RabMAb|Recombinant|KO Validated|20ul selling size
![Immunocytochemistry/ Immunofluorescence - Anti-MLH1 antibody [EPR3894] (AB92312)](https://content.abcam.com/products/images/mlh1-antibody-epr3894-ab92312--immunocytochemistry-immunofluorescence-img13107.jpg)
- 5
- 4
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 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 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: 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