Why is my protein sample aggregating?

1. Hydrophobic Interactions: Proteins have hydrophobic regions that tend to interact with each other, leading to aggregation when exposed to an aqueous environment.
2. pH Variations: Changes in pH can disrupt the electrostatic interactions between proteins, causing them to aggregate.
3. Ionic Strength: Variations in salt concentration can affect protein solubility and lead to aggregation.
4. Temperature: High temperatures can denature proteins and promote their aggregation.
5. Concentration: Extremely high protein concentrations can exceed the saturation point and trigger aggregation.
6. Shear Stress: Mechanical stress, such as pipetting or vortexing, can disrupt protein structure and lead to aggregation.
7. Protein Purity: Contaminants or impurities in protein samples can promote aggregation.
8. Presence of Aggregating Agents: Some molecules, like detergents or denaturing agents, can induce protein aggregation.
9. Protein Modifications: Post-translational modifications or chemical modifications can alter protein properties and lead to aggregation.
10. Protein Size: Larger proteins are more prone to aggregation due to increased molecular interactions.
11. Protein Structure: Proteins with inherently unstable or misfolded structures are more likely to aggregate.
12. Time: Prolonged storage or incubation can promote aggregation as proteins may gradually lose stability.
13. Protein-Protein Interactions: Interactions between multiple proteins can result in the formation of aggregates or complexes.
14. Deterioration of Sample Conditions: Improper storage conditions, such as freeze-thaw cycles or exposure to light, can lead to sample degradation and aggregation.

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