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Rabbit Recombinant Monoclonal MLH1 antibody. Suitable for WB, IHC-P, ICC/IF, Flow Cyt (Intra) and reacts with Mouse, Rat, Human samples. Cited in 83 publications.


Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLH1 antibody [EPR3894] (AB92312), expandable thumbnail
  • Western blot - Anti-MLH1 antibody [EPR3894] (AB92312), expandable thumbnail
  • Western blot - Anti-MLH1 antibody [EPR3894] (AB92312), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-MLH1 antibody [EPR3894] (AB92312), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLH1 antibody [EPR3894] (AB92312), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWBIHC-PICC/IFFlow Cyt (Intra)
Human
Not recommended
Tested
Tested
Tested
Tested
Mouse
Not recommended
Tested
Expected
Expected
Expected
Rat
Not recommended
Tested
Expected
Expected
Expected

Not recommended
Not recommended

Species

Mouse, Rat, Human

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/2000

Notes

For unpurifid use at 1/10000 - 1/50000

Species

Rat

Dilution info

1/2000

Notes

For unpurifid use at 1/10000 - 1/50000

Species

Human

Dilution info

1/2000

Notes

For unpurifid use at 1/10000 - 1/50000

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/500

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/10 - 1/100

Notes

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

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Target data

Function

Heterodimerizes with PMS2 to form MutL alpha, a component of the post-replicative DNA mismatch repair system (MMR). DNA repair is initiated by MutS alpha (MSH2-MSH6) or MutS beta (MSH2-MSH3) binding to a dsDNA mismatch, then MutL alpha is recruited to the heteroduplex. Assembly of the MutL-MutS-heteroduplex ternary complex in presence of RFC and PCNA is sufficient to activate endonuclease activity of PMS2. It introduces single-strand breaks near the mismatch and thus generates new entry points for the exonuclease EXO1 to degrade the strand containing the mismatch. DNA methylation would prevent cleavage and therefore assure that only the newly mutated DNA strand is going to be corrected. MutL alpha (MLH1-PMS2) interacts physically with the clamp loader subunits of DNA polymerase III, suggesting that it may play a role to recruit the DNA polymerase III to the site of the MMR. Also implicated in DNA damage signaling, a process which induces cell cycle arrest and can lead to apoptosis in case of major DNA damages. Heterodimerizes with MLH3 to form MutL gamma which plays a role in meiosis.

Alternative names

DNA mismatch repair protein Mlh1, MutL protein homolog 1, COCA2, MLH1

Recommended products

Rabbit Recombinant Monoclonal MLH1 antibody. Suitable for WB, IHC-P, ICC/IF, Flow Cyt (Intra) and reacts with Mouse, Rat, Human samples. Cited in 83 publications.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR3894

Purification technique

Affinity purification Protein A

Specificity

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

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

Activity summary

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.

Associated diseases and disorders

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

15 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLH1 antibody [EPR3894] (ab92312), expandable thumbnail
    Image from Wang X et al. PLoS One. 2011;6(10):e25913. Epub 2011 Oct 12. Fig 3.; doi:10.1371/journal.pone.0025913; October 12 2011 PLoS ONE 6(10): e25913.

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLH1 antibody [EPR3894] (ab92312)

    Unpurified ab92312 staining MLH1 in Human colorectal (top) and gastric tissue (bottom) by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

  • Western blot - Anti-MLH1 antibody [EPR3894] (ab92312), expandable thumbnail

    Western blot - Anti-MLH1 antibody [EPR3894] (ab92312)

    False colour image of Western blot: Anti-MLH1 antibody [EPR3894] staining at 1/2000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control 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 Human MLH1 knockout A549 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°C. 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.

    All lanes: Western blot - Anti-MLH1 antibody [EPR3894] (ab92312) 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 3: Jurkat cell lysate at 20 µg

    Lane 4: HCT 116 cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 85 kDa

    Observed band size: 85 kDa

  • Western blot - Anti-MLH1 antibody [EPR3894] (ab92312), expandable thumbnail

    Western blot - Anti-MLH1 antibody [EPR3894] (ab92312)

    Lanes 1-4: Merged signal (red and green). Green - ab92312 observed at 90 kDa. Red - loading control Anti-GAPDH antibody [6C5] - 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 Human MLH1 knockout HeLa cell line ab267223 (knockout cell lysate Human MLH1 knockout HeLa 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 (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 20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-MLH1 antibody [EPR3894] (ab92312) 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 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size: 85 kDa

    Observed band size: 85 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-MLH1 antibody [EPR3894] (ab92312), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-MLH1 antibody [EPR3894] (ab92312)

    Immunocytochemistry/ Immunofluorescence analysis of SW480 (Human colorectal adenocarcinoma epithelial cell) cells labeling MLH1 with Purified ab92312 at 1:500 dilution (1.6 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor®594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor®488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLH1 antibody [EPR3894] (ab92312), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLH1 antibody [EPR3894] (ab92312)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon tissue sections labeling MLH1 with Purified ab92312 at 1:250 dilution (2.9 μg/ml). Heat mediated antigen retrieval was performed using citrate (pH 6.0)ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody.Negative control:PBS instead of the primary antibody.Hematoxylinwas used as a counterstain

  • Western blot - Anti-MLH1 antibody [EPR3894] (ab92312), expandable thumbnail

    Western blot - Anti-MLH1 antibody [EPR3894] (ab92312)

    Blocking and diluting buffer: 5% NFDM/TBST.
    According to PMID:23653048, HCT116 is MLH1 negative cell line.

    All lanes: Western blot - Anti-MLH1 antibody [EPR3894] (ab92312) at 0.4 µg/mL

    Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg

    Lane 2: HCT116 (Human colorectal carcinoma epithelial cell) whole cell lysates, negative control at 20 µg

    Lane 3: Human testis lysates at 20 µg

    Lane 4: Rat testis lysates at 20 µg

    Lane 5: Mouse thymus lysates at 20 µg

    Lane 6: Rat thymus lysates at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 85 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-MLH1 antibody [EPR3894] (ab92312), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-MLH1 antibody [EPR3894] (ab92312)

    Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) labelling MLH1 with purified ab92312 at 1/1000. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077). Nuclei counterstained with DAPI (blue).

    Control: PBS only

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLH1 antibody [EPR3894] (ab92312), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLH1 antibody [EPR3894] (ab92312)

    Unpurified ab92312 at 1/100 dilution staining MLH1 in Human colonic adenocarcinoma by Immunohistochemistry, Paraffin-embedded tissue. The use of an HRP/AP polymerized antibody is recommended for a secondary antibody. Heat mediated antigen retrieval was performed via the pressure cooker method before commencing with IHC staining protocol.

  • Western blot - Anti-MLH1 antibody [EPR3894] (ab92312), expandable thumbnail

    Western blot - Anti-MLH1 antibody [EPR3894] (ab92312)

    All lanes: Western blot - Anti-MLH1 antibody [EPR3894] (ab92312) at 1/10000 dilution

    Lane 1: 293 cell lysate at 10 µg

    Lane 2: HeLa cell lysate at 10 µg

    Lane 3: A431 cell lysate at 10 µg

    Lane 4: SW480 cell lysate at 10 µg

    Secondary

    All lanes: goat anti-rabbit HRP at 1/2000 dilution

    Predicted band size: 85 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-MLH1 antibody [EPR3894] (ab92312), expandable thumbnail
    This image is courtesy of a customer review submitted by Kirk McManus.

    Immunocytochemistry/ Immunofluorescence - Anti-MLH1 antibody [EPR3894] (ab92312)

    Unpurified ab92312 (1/200) staining MLH1 in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X100/PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to abreview.

  • Flow Cytometry (Intracellular) - Anti-MLH1 antibody [EPR3894] (ab92312), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-MLH1 antibody [EPR3894] (ab92312)

    Overlay histogram showing HeLa cells stained with unpurifiedab92312 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab92312, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLH1 antibody [EPR3894] (ab92312), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLH1 antibody [EPR3894] (ab92312)

    Unpurified ab92312 at 1/100 dilution staining MLH1 in Human tonsil by Immunohistochemistry, Paraffin-embedded tissue. The use of an HRP/AP polymerized antibody is recommended for a secondary antibody. Heat mediated antigen retrieval was performed via the pressure cooker method before commencing with IHC staining protocol.

  • Western blot - Anti-MLH1 antibody [EPR3894] (ab92312), expandable thumbnail

    Western blot - Anti-MLH1 antibody [EPR3894] (ab92312)

    Lanes 1 - 4: Merged signal (red and green). Green - ab92312 observed at 88 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.


    Unpurified ab92312 was shown to recognize MLH1 in wild-type HAP1 cells along with additonal cross reactive bands. No band was observed when MLH1 knockout samples were examined. Wild-type and MLH1 knockout samples were subjected to SDS-PAGE. ab92312 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (loading control to GAPDH) were both diluted 1/1000 and incubated overnight at 4°C. 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 1/10,000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-MLH1 antibody [EPR3894] (ab92312) at 1/1000 dilution

    Lane 1: Wild-type HAP1 cell lysate at 20 µg

    Lane 2: MLH1 knockout HAP1 cell lysate at 20 µg

    Lane 3: HCT116 cell lysate at 20 µg

    Lane 4: 293T cell lysate at 20 µg

    Predicted band size: 85 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLH1 antibody [EPR3894] (ab92312), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLH1 antibody [EPR3894] (ab92312)

    Tissue Microarrays stained for "Anti-MLH1 antibody [EPR3894]" using "ab92312"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Citrate Buffer) ab93678 (citrate buffer, pH 6.0). The sections were incubated with ab92312 at +4°C overnight. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody.

  • Flow Cytometry (Intracellular) - Anti-MLH1 antibody [EPR3894] (ab92312), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-MLH1 antibody [EPR3894] (ab92312)

    Flow cytometry overlay histogram showing wild-type Hap1 (green line) and MLH1 knockout Hap1 stained with ab92312 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab92312) (1x 106 in 100μl at 0.2 μg/ml (1/9835)) for 30min at 22°C.

    The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed was incubated at 1/4000 for 30min at 22°C

    Isotype control antibody Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control was used at the same concentration and conditions as the primary antibody (wild-type Hap1 - black line, MLH1 knockout Hap1 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

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