United Kingdom (UK)
Recombinant monoclonal antibody to MSH2. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma 1B11-14-16.
UniProt Accession Number of Target Protein: P43246
Alternative Name(s) of Target: hMSH2; DNA mismatch repair protein Msh2; MutS protein homolog 2; CGMCC NO.17405; G219-1129
Immunogen: The original antibody was generated by immunizing BALB/c mice with human MSH2 protein peptide (KLH-NSFVNEIISRIKVTT-Cys) conjugated with KLH in equal volume of Freund's complete adjuvant.
Specificity: This antibody binds an epitope on the human DNA mismatch protein MSH2 between amino acids 919-934 (NSFVNEIISRIKVTT). MSH2 forms a heterodimer with MSH6 to make the human MutSα mismatch repair complex. The DNA mismatch repair system (MMR) is composed of enzyme molecules that specifically repair DNA base mismatches. MSH2 is involved in many different forms of DNA repair, including transcription-coupled repair, homologous recombination, and base excision repair. Studies have shown that mutations and/or loss of function of MMR genes are related to the occurrence of a variety of tumors, such as colorectal cancer, skin cancer, ovarian cancer, and endometrial cancer.
Application Notes: The binding specificity of this antibody for human MSH2 was determined using ELISA. This antibody was capable of detecting MSH2 from HCT8 and Biu-87 cell lysates in a western blot. This antibody was also used in the immunohistochemical analysis of normal tissues like tonsil, appendix tissues and cancerous tissues like colorectal and bladder cancer. This antibody shows similar staining of MSH2 protein compared to the commercially available antibody G219-1129 (CN110218251).
Antibody first published in: PMID:
Flow cytometry using anti-MSH2 antibody 1B11-14-16 (Ab03191). Paraformaldehyde fixed HeLa cells permeabilized with 0.5% Triton were stained with the anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG version of 1B11-14-16 (Ab03191-23.0, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-rabbit IgG AlexaFluor® 488 antibody at a dilution of 1:1000, and the cells were analyzed using a FACSCanto flow-cytometer.
Western blot using anti-MSH2 antibody 1B11-14-16 (Ab03191). HEK293 (A) (0.01µg/ml), A431 (B) (0.01µg/ml), HeLa (C) (0.01µg/ml), HepG2 (D) (0.03µg/ml), K562 (E) (0.03µg/ml) cells nuclear lysate, and A549 (F) (0.03µg/ml) cells lysate (35µg protein in RIPA buffer) were resolved on an SDS-PAGE gel, and blots were probed with the chimeric rabbit version of 1B11-14-16 (Ab03191-23.0) at the mentioned respective concentrations before detection using an anti-rabbit secondary antibody. A primary incubation of 1 hour was used, and protein was detected by chemiluminescence.
Immunofluorescence staining of HeLa cells with anti-MSH2 antibody 1B11-14-16 (Ab03191). Immunofluorescence analysis of paraformaldehyde fixed HeLa cells, permeabilized with 0.15% Triton, on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of 1B11-14-16 (Ab03191-23.0) (1:100 dilution) for 1h followed by Alexa Fluor® 488 secondary antibody (1:1000 dilution), showing cytoplasmic and some nuclear staining. The nuclear stain is DAPI (blue). Panels show, from left-right, top-bottom, Ab03191-23.0, DAPI, merged channels, and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.