Product nameMismatch Repair (MSH6, PMS2, MLH1, MSH2) Antibody Panel - Human
Species reactivityReacts with: Human
Mismatch Repair (MSH6, PMS2, MLH1, MSH2) Antibody Panel - Human ab252190 contains multiple trial-sized versions of anti-human antibody clones against MSH6, PMS2, MLH1, and MSH2, specifically selected for their high performance in multiple applications including IHC. This panel contains 4 recombinant rabbit monoclonal antibodies that are all knock-out validated to ensure specificity to their targets. They are provided as a sampler panel to allow you to easily evaluate each in your required application.
DNA mismatch repair (MMR) proteins are involved in repairing mistakes that occur during DNA replication and recombination, in addition to repairing some types of DNA damage. Defects in the MMR process due to mutations in MMR genes (MSH6, PMS2, MLH1, MSH2) can result in microsatellite instability (MSI), where a DNA sequence accumulates errors and produces abnormally long or shorter microsatellites. These defects in the MMR pathway have been linked to various human cancers, such as human non-polyposis colon cancer (HNPCC) and Muir-Torre Syndrome (MTS), a subtype of HNPCC.
For guidelines on how to use each antibody within the panel, please consult the individual datasheet for each antibody.
- Rabbit monoclonal [EPR3894] to MLH1 (20 µL) ab92312
- Rabbit monoclonal [EPR21017-123] to MSH2 (20 µL) ab227941
- Rabbit monoclonal [EPR3945] to MSH6 (20 µL) ab92471
- Rabbit monoclonal [EPR3947] to PMS2 (20 µL) ab110638
Explore our range of antibody sample panels designed to provide you with a variety of trial-size antibodies in a convenient and cost-effective format.
Carrier-free formulations of our recombinant antibodies are available and ready to use for multiplex IHC analysis including Imaging Mass CytometryTM. Please refer to the ‘Associated products’ section below.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Storage instructionsStore at -20°C. Please refer to protocols.
Components 1 kit ab92312 - Anti-MLH1 antibody [EPR3894] 2 x 10µl ab227941 - Anti-MSH2 antibody [EPR21017-123] 2 x 10µl ab92471 - Anti-MSH6 antibody [EPR3945] 2 x 10µl ab110638 - Anti-PMS2 antibody [EPR3947] 2 x 10µl
Cellular localizationMSH2: Nucleus. PMS2: Nucleus. MLH1: Nucleus. MSH6: Nucleus.
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
- Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
- Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777)
Immunohistochemical staining of paraffin embedded human colon with purified ab92471 at a dilution of 1/500. A pre-diluted HRP polymer for rabbit/mouse IgG was used as the secondary antibody and the sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
ab92471 staining MSH6 in wild-type HAP1 cells (top panel) and MSH6 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab92471 at 1μg/ml and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
ab110638 at 1/100 dilution staining PMS2 in Human colonic adenocarcinoma by Immunohistochemistry, Paraffin-embedded tissue.
Lane 1: Wild-type HAP1 whole cell lysate (30 µg)
Lane 2: PMS2 knockout HAP1 whole cell lysate (30 µg)
Lanes 1-2: Merged signal (red and green). Green - Anti-PMS2 antibody [EPR3947] (ab110638) observed at 120 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab110638 was shown to specifically react with PMS2 in wild-type HAP1 cells. No band was observed when PMS2 knockout samples were used. Wild-type and PMS2 knockout samples were subjected to SDS-PAGE. Ab110638 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling MSH2 with ab227941 at 1/8000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining in para-carcinoma colonic epithelium (image B) or stromal cells (both image A and B) and loss of expression in the paired colon cancer (image A) (PMID: 24710284). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Immunofluorescent analysis of 100% methanol-fixed A549 (human lung carcinoma cell line) cells labeling MSH2 with ab227941 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Nuclear staining in A549 cell line is shown.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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 Perform heat mediated antigen retrieval 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.
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: MLH1 knockout HAP1 cell lysate (20 µg)
Lane 3: HCT116 cell lysate (20 µg)
Lane 4: 293T cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - Anti-MLH1 antibody Anti-MLH1 antibody [EPR3894] (ab92312) observed at 88 kDa. Red - 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 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 (ab216773) and 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.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab252190 has been referenced in 1 publication.
- Tian T et al. Microsatellite instability and its associations with the clinicopathologic characteristics of diffuse large B-cell lymphoma. Cancer Med 9:2330-2342 (2020). PubMed: 32022486