Applications of DL-Dithiothreitol (DTT)  

DL-Dithiothreitol (DTT) is a strong reducing agent that breaks disulfide bonds (—S—S—) in proteins and other biomolecules. It is widely used in biochemistry, molecular biology, and biotechnology for stabilizing and modifying biomolecules.  

1. Protein Research & Analysis  

Disulfide Bond Reduction  

- Denaturing proteins for SDS-PAGE, Western blotting, or mass spectrometry by linearizing proteins.  

-Preventing protein aggregation during extraction and purification.  

- Refolding recombinant proteins (e.g., from inclusion bodies) by controlling disulfide bond formation.  

Enzyme Activity Preservation

- Maintains thiol-dependent enzymes (e.g., dehydrogenases, proteases) in their active state by keeping cysteine residues reduced.  

Antibody Processing 

- Generates Fab fragments by reducing disulfide bonds in antibodies (e.g., for immunoassays).  

2. Nucleic Acid Research 

RNA Protection  

- Prevents RNA degradation by inhibiting RNases (often used with RNase inhibitors).  

- Used in RT-PCR, RNA extraction, and cDNA synthesis** protocols.  

DNA Polymerase Stability 

- Some DNA polymerases (e.g., Taq) require DTT to maintain activity by protecting essential thiol groups.  

3. Cell Biology & Biochemistry

Cell Lysis & Protein Extraction 

- Added to lysis buffers (e.g., RIPA) to prevent protein oxidation.  

Studying Membrane Proteins  

- Used to investigate disulfide bonds in receptors or ion channels.  

4. Biopharmaceutical Applications 

- Protein drug formulation (prevents unwanted disulfide bond formation).  

- Recombinant protein production (ensures proper folding).