MLH1 Antibody (SP08-04)

Western Blot: MLH1 Antibody (SP08-04) [NBP2-67381] - Western blot analysis of MLH1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabb

Immunocytochemistry/Immunofluorescence: MLH1 Antibody (SP08-04) [NBP2-67381] - Staining MLH1 in HepG2 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

Immunohistochemistry-Paraffin: MLH1 Antibody (SP08-04) [NBP2-67381] - Immunohistochemical analysis of paraffin-embedded rat large intestine tissue using anti-MLH1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1604-41, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Flow Cytometry: MLH1 Antibody (SP08-04) [NBP2-67381] - Analysis of Hela cells with MLH1 antibody at 1/50 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody

Immunohistochemistry-Paraffin: MLH1 Antibody (SP08-04) [NBP2-67381] - Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-MLH1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1604-41, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemistry-Paraffin: MLH1 Antibody (SP08-04) [NBP2-67381] - Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-MLH1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1604-41, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Knockout Validated: MLH1 Antibody (SP08-04) [NBP2-67381] - All lanes: Western blot analysis of MLH1 with anti-MLH1 antibody [SP08-04] at 1:1,000 dilution. Lane 1: Wild-type B16F10 whole cell lysate. Lane 2: MLH1 knockout B16F10 whole cell lysate. NBP2-67381 was shown to specifically react with MLH1 in wild-type B16F10 cells. No band was observed when MLH1 knockout samples were tested. Wild-type and MLH1 knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary Anti-MLH1 antibody (1/1,000) and Anti-Vinculin antibody (1/5,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody at 1:200,000 dilution was used for 1 hour at room temperature.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Flow, ICC/IF, IHC, IHC-P, IP, KO
• Clone: SP08-04
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

MLH1 Antibody (SP08-04) Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: Synthetic peptide within Human MLH1 aa 707-756 / 756. (SwissProt: P40692 Human; SwissProt: Q9JK91 Mouse; SwissProt: P97679 Rat)
• Localization: Nucleus.
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: MLH1
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:50-1:100
  • Immunocytochemistry/Immunofluorescence 1:50-1:200
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:50-1:200
  • Immunoprecipitation
  • Knockout Validated
  • Western Blot 1:500-1:1000

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: TBS (pH7.4), 0.05% BSA, 40% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for MLH1 Antibody (SP08-04)

• COCA2FCC2
• DNA mismatch repair protein Mlh1
• hMLH1
• HNPCC
• HNPCC2MGC5172
• mutL (E. coli) homolog 1 (colon cancer, nonpolyposis type 2)
• mutL homolog 1, colon cancer, nonpolyposis type 2 (E. coli)
• MutL protein homolog 1

Background

MLH1 is a DNA mismatch repair protein. The repair of mismatch DNA is essential to maintaining the integrity of genetic information over time. An alteration of microsatellite repeats is the result of slippage owing to strand misalignment during DNA replication and is referred to as microsatellite instability (MSI). These defects in DNA repair pathways have been related to human carcinogenesis. The importance of mismatch repair genes became apparent with the identification of the genetic basis for hereditary nonpolyposis colon cancer (HNPCC). MSHS2 is involved in the initial cognition of mismatch nucleotides during the replication mismatch repair process. It is thought that after MSH2 binds to a mismatched DNA duplex it is joined by a heterodimer of MLH1 and PMSH, which together help facilitate the later steps in mismatch repair.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: MLH1