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AB257172

Human MLH1 knockout HeLa cell lysate

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MLH1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon12 and 1 bp deletion in exon12 and 4 bp deletion in exon12.
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Sanger Sequencing - Human MLH1 knockout HeLa cell lysate (AB257172)
  • Sanger seq

Unknown

Sanger Sequencing - Human MLH1 knockout HeLa cell lysate (AB257172)

Allele-2 : 4 bp deletion in exon12

Sanger Sequencing - Human MLH1 knockout HeLa cell lysate (AB257172)
  • Sanger seq

Unknown

Sanger Sequencing - Human MLH1 knockout HeLa cell lysate (AB257172)

Allele-1 : 10 bp deletion in exon12

Sanger Sequencing - Human MLH1 knockout HeLa cell lysate (AB257172)
  • Sanger seq

Unknown

Sanger Sequencing - Human MLH1 knockout HeLa cell lysate (AB257172)

Allele-3 : 1 bp deletion in exon12

Western blot - Human MLH1 knockout HeLa cell lysate (AB257172)
  • WB

Supplier Data

Western blot - Human MLH1 knockout HeLa cell lysate (AB257172)

Lane 1 : Wild-type HeLa cell lysate (20 μg)

Lane 2 : MLH1 knockout HeLa cell lysate (20 μg)

Lane 3 : Jurkat cell lysate (20 μg)

Lane 4 : HCT116 cell lysate (20 μg)

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°C 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:

Western blot - Human MLH1 knockout HeLa cell lysate (ab257172) at 20 µg

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

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Western blot - Human MLH1 knockout HeLa cell lysate (AB257172)
  • WB

Supplier Data

Western blot - Human MLH1 knockout HeLa cell lysate (AB257172)

Lane 1 : Wild-type HeLa cell lysate (20 μg)

Lane 2 : MLH1 knockout HeLa cell lysate (20 μg)

Lane 3 : Jurkat cell lysate (20 μg)

Lane 4 : HCT116 cell lysate (20 μg)

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°C 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:

Western blot - Human MLH1 knockout HeLa cell lysate (ab257172) at 20 µg

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

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon12 and 1 bp deletion in exon12 and 4 bp deletion in exon12.

Disease

Adenocarcinoma

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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

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What's included?

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Properties and storage information

Gene name
MLH1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Quality control - STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

MLH1 also known as MutL homolog 1 is a protein involved in DNA mismatch repair an important mechanism for maintaining genetic stability. It has a molecular weight of approximately 87 kDa. This protein is expressed in various tissues but is most abundant in the colonic epithelium and endometrium. MLH1 acts mechanically by forming heterodimers with other proteins collaborating in correcting errors that occur during DNA replication.
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.

MLH1 mutations are closely linked to Lynch syndrome and sporadic colorectal cancer. Lynch syndrome a hereditary condition significantly raises the risk of colorectal cancer and other cancers due to defective DNA mismatch repair. MLH1 mutations often lead to the loss of MLH1 protein expression particularly observed in MLH1 IHC staining. Additionally in colorectal cancer the MLH1 protein may interact with APC and TP53 playing a role in cancer progression and tumorigenesis.
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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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