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     Black line: Control Antigen (100 ng); Purple line: Antigen(10ng); Blue line: Antigen (50 ng); Red line: Antigen (100 ng);    

  
  

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     Western blot analysis using MGMT mouse mAb against MCF-7 (1), Jurkat (2), HepG2 (3), and PC-3 (4) cell lysate.    

  
  

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     Immunofluorescence analysis of Hela cells using MGMT mouse mAb (green). Blue: DRAQ5 fluorescent DNA dye. Red: Actin filaments have been labeled with Alexa Fluor- 555 phalloidin. Secondary antibody from Fisher (Cat#: 35503)    

  
  

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     Flow cytometric analysis of Hela cells using MGMT mouse mAb (green) and negative control (red).    

  
  

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     Immunohistochemical analysis of paraffin-embedded human rectum cancer tissues using MGMT mouse mAb with DAB staining.    

  
  

MGMT (O?-methylguanine-DNA methyltransferase) is a DNA repair enzyme that plays a critical role in maintaining genomic integrity by removing alkyl groups from the O? position of guanine, a common site of DNA damage induced by alkylating chemotherapeutic agents like temozolomide. In cancer biology, MGMT expression is inversely associated with treatment response; high MGMT activity can counteract the cytotoxic effects of alkylating drugs by repairing DNA damage, thereby conferring tumor resistance.

Epigenetic silencing of the *MGMT* gene via promoter methylation is a key mechanism leading to reduced enzyme expression. This methylation status has emerged as a predictive biomarker in glioblastoma (GBM), where methylated *MGMT* promoters correlate with improved survival in patients treated with temozolomide and radiotherapy.

MGMT antibodies are widely used in immunohistochemistry (IHC) to detect protein expression in tumor tissues. However, IHC results may not always align with methylation assays (e.g., methylation-specific PCR or pyrosequencing) due to post-transcriptional regulation or technical variability. Despite this, IHC remains a practical tool for assessing MGMT protein levels in resource-limited settings.

Clinical guidelines increasingly prioritize molecular testing for methylation status, but MGMT antibodies retain utility in research and diagnostic workflows to explore protein expression patterns and validate epigenetic findings. Their application underscores the importance of MGMT as a therapeutic biomarker in tailoring alkylating chemotherapy for cancers, particularly gliomas.