Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-PMS2 antibody [EPR3947] (ab110638)

Knockout Tested Rabbit recombinant monoclonal PMS2 antibody [EPR3947]. Validated in WB, IP, IHC, Flow Cyt, ICC/IF and tested in Human. Cited in 7 publication(s). Independently reviewed in 2 review(s).

Overview

  • Product name

    Anti-PMS2 antibody [EPR3947]
    See all PMS2 primary antibodies
  • Description

    Rabbit monoclonal [EPR3947] to PMS2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human PMS2 aa 1-100. The exact sequence is proprietary.

  • Positive control

    • WB: Wild-type HAP1 cell lysate; Jurkat, HeLa, SH-SY5Y and SKBR3 cell lysates. IHC-P: Human colon and colon adenocarcinoma tissue. ICC/IF: HeLa and Raji cells. Flow cyt: HeLa cells.
  • General notes

    To see more of the key markers and tools you need to study the hallmarks of cancer, including genome instability and mutation, please visit the following page.

    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 rabbit monoclonal antibody.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Dissociation constant (KD)

    KD = 1.50 x 10 -10 M
    Learn more about KD
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol, 0.05% BSA, 50% Tissue culture supernatant
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EPR3947
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab110638 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000 - 1/10000. Detects a band of approximately 110 kDa (predicted molecular weight: 96 kDa).
IP 1/10 - 1/100.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF 1/100 - 1/250.
Flow Cyt 1/1000.

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

 

Target

  • Function

    Component of the post-replicative DNA mismatch repair system (MMR). Heterodimerizes with MLH1 to form MutL alpha. DNA repair is initiated by MutS alpha (MSH2-MSH6) or MutS beta (MSH2-MSH6) 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. MulL 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.
  • Involvement in disease

    Defects in PMS2 are the cause of hereditary non-polyposis colorectal cancer type 4 (HNPCC4) [MIM:600259]. Mutations in more than one gene locus can be involved alone or in combination in the production of the HNPCC phenotype (also called Lynch syndrome). Most families with clinically recognized HNPCC have mutations in either MLH1 or MSH2 genes. HNPCC is an autosomal, dominantly inherited disease associated with marked increase in cancer susceptibility. It is characterized by a familial predisposition to early onset colorectal carcinoma (CRC) and extra-colonic cancers of the gastrointestinal, urological and female reproductive tracts. HNPCC is reported to be the most common form of inherited colorectal cancer in the Western world, and accounts for 15% of all colon cancers. Cancers in HNPCC originate within benign neoplastic polyps termed adenomas. Clinically, HNPCC is often divided into two subgroups. Type I: hereditary predisposition to colorectal cancer, a young age of onset, and carcinoma observed in the proximal colon. Type II: patients have an increased risk for cancers in certain tissues such as the uterus, ovary, breast, stomach, small intestine, skin, and larynx in addition to the colon. Diagnosis of classical HNPCC is based on the Amsterdam criteria: 3 or more relatives affected by colorectal cancer, one a first degree relative of the other two; 2 or more generation affected; 1 or more colorectal cancers presenting before 50 years of age; exclusion of hereditary polyposis syndromes. The term 'suspected HNPCC' or 'incomplete HNPCC' can be used to describe families who do not or only partially fulfill the Amsterdam criteria, but in whom a genetic basis for colon cancer is strongly suspected.
    Defects in PMS2 are a cause of mismatch repair cancer syndrome (MMRCS) [MIM:276300]; also known as Turcot syndrome or brain tumor-polyposis syndrome 1 (BTPS1). MMRCS is an autosomal dominant disorder characterized by malignant tumors of the brain associated with multiple colorectal adenomas. Skin features include sebaceous cysts, hyperpigmented and cafe au lait spots.
  • Sequence similarities

    Belongs to the DNA mismatch repair mutL/hexB family.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • DNA mismatch repair gene homologue antibody
    • DNA mismatch repair protein PMS2 antibody
    • H_DJ0042M02.9 antibody
    • HNPCC4 antibody
    • Mismatch repair endonuclease PMS2 antibody
    • Mismatch repair gene PMSL2 antibody
    • MLH4 antibody
    • PMS 2 antibody
    • PMS1 homolog 2 mismatch repair system antibody
    • PMS1 protein homolog 2 antibody
    • PMS2 antibody
    • PMS2 postmeiotic segregation increased 2 antibody
    • PMS2 postmeiotic segregation increased 2 (S. cerevisiae) antibody
    • PMS2_HUMAN antibody
    • PMS2CL antibody
    • PMSL2 antibody
    • Postmeiotic segregation increased, S. cerevisiae, 2 antibody
    see all

Images

  • 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 - 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.

  • Immunofluorescent analysis of 0.1% TritonX-100-fixed, DAPI permeabilized Raji (human burkitt's lymphoma b lymphocyte) cells labelling PMS2 with ab110638 at 1/500 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in Raji cells. 4% Paraformaldehyde was used to counterstain tubulin at  dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

  • Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling PMS2 with ab110638, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on human colon. The section was incubated with ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

  • Anti-PMS2 antibody [EPR3947] (ab110638) at 1/1500 dilution + HeLa (Human cervix adenocarcinoma epithelial cell) Whole cell lysates at 15 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 96 kDa
    Observed band size: 110 kDa
    why is the actual band size different from the predicted?



    Blocking/diluting buffer and concentration: 5% NFDM/TBST.

    Exposure time: 50 seconds

  • Overlay histogram showing HeLa cells stained with ab110638 (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 (ab110638, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
  • ab110638 at 1/100 dilution staining PMS2 in Human colonic adenocarcinoma by Immunohistochemistry, Paraffin-embedded tissue.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • ab110638 at 1/100 dilution staining PMS2 in HeLa cells by Immunofluorescence.
  • Immunocytochemistry/ Immunofluorescence analysis of HeLa cell labeling PMS2 with ab110638 at 1/400 dilution. Goat anti-Rabbit Alexa fluor®488 at 1/200 was used as the secondary antibody. Cells were fixed with Paraformaldehyde and permeabilised with 0.5% Triton-X100 in PBS.

     

    See Abreview

  • All lanes : Anti-PMS2 antibody [EPR3947] (ab110638) at 1/1000 dilution

    Lane 1 : Jurkat cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : SH SY5Y cell lysate
    Lane 4 : SKBR3 cell lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 96 kDa
    Observed band size: 110 kDa why is the actual band size different from the predicted?

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References

This product has been referenced in:

  • Aasebø KØ  et al. Consequences of a high incidence of microsatellite instability and BRAF-mutated tumors: A population-based cohort of metastatic colorectal cancer patients. Cancer Med 8:3623-3635 (2019). Read more (PubMed: 31070306) »
  • Zhou ZH  et al. The prognostic value and pathobiological significance of Glasgow microenvironment score in gastric cancer. J Cancer Res Clin Oncol N/A:N/A (2017). IHC ; Human . Read more (PubMed: 28180998) »
See all 8 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton-X100 in PBS

Dr. Kirk Mcmanus

Verified customer

Submitted Apr 20 2012

Application
Western blot
Sample
Human Cell lysate - whole cell (different cell lines)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
30 µg
Specification
different cell lines
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Pounami Samadder

Verified customer

Submitted Mar 22 2019

Application
Western blot
Sample
Human Cell lysate - whole cell (HCT116 cell lines)
Gel Running Conditions
Reduced Denaturing (8)
Loading amount
30 µg
Treatment
Expression seen with MLH1 expression
Specification
HCT116 cell lines
Blocking step
odyssey PBS as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: 37°C

Abcam user community

Verified customer

Submitted Jul 14 2015

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