A radioisotope is an isotope of an element with an unstable nucleus, causing it to emit alpha, beta, or gamma rays as it decays to a more stable form. Radioisotopes are used in biochemistry for tracing molecular pathways, studying enzyme kinetics, labelling biomolecules, and conducting sensitive diagnostic assays due to their ability to be easily detected through the radiation they emit.
Common Radioisotopes in Biochemistry:
- Carbon-14 (⁶C¹⁴)
- Phosphorus-32 (¹⁵P³²)
- Tritium (Hydrogen-3) (¹H³)
- Sulfur-35 (¹⁶S³⁵)
- Iodine-125 (⁵³I¹²⁵)
Uses of Radioisotopes in Biochemistry:
- Tracer Studies:
- Carbon-14 and Phosphorus-32 are used in metabolic studies to track how molecules move through biochemical pathways. For instance, they can trace the fate of glucose, fatty acids, or amino acids in metabolic reactions, allowing researchers to understand how cells utilize these molecules.
- Radioimmunoassays (RIA):
- Iodine-125 is used in RIA to quantify hormones, antigens, and other biological molecules in the blood. This highly sensitive technique is essential for clinical diagnostics, as it can detect minute concentrations of substances like insulin, cortisol, or thyroid hormones.
- DNA/RNA Research:
- Phosphorus-32 and Tritium label nucleotides for studying DNA replication, transcription, and gene expression. This helps detect DNA or RNA sequences through Southern or Northern blotting.
- Protein Labelling:
- Sulfur-35 and Iodine-125 label proteins in protein structure studies, enzyme-substrate interactions, and electrophoresis. This is particularly useful in SDS-PAGE for protein separation and identification.
- Enzyme Kinetics and Phosphorylation Studies:
- Phosphorus-32 is used to study phosphorylation, a key regulatory process in biochemistry where phosphate groups are added to proteins. This isotope allows researchers to observe how enzymes like kinases transfer phosphate groups in cell signalling pathways.
- Autoradiography:
- This technique involves using radioisotopes (e.g., Tritium) to visualize the location of a labelled molecule in a biological sample. It is particularly useful in visualizing DNA or RNA distribution in tissues or cells.
- Medical Biochemistry:
- In clinical biochemistry labs, radioisotopes like Iodine-125 are used in assays to detect hormone-related disorders, such as hypothyroidism or hyperthyroidism.
Safety Considerations in Biochemistry Labs:
Working with radioisotopes requires stringent safety protocols, including:
- Radiation shielding (lead-lined equipment)
- Personal dosimeters to monitor radiation exposure
- Proper waste disposal procedures for radioactive materials
- Controlled environments to prevent contamination