15 microliters of sample and 50 microliters of hydrogen peroxide. how do I modify these volumes to avoid too much bubbles
The cause of excessive bubbling was as follows:
Catalase Activity: Fresh liver tissues contain high concentrations of catalase, an enzyme that accelerates H₂O₂ decomposition into water and oxygen gas (bubbles). Your current ratio (15 µL sample : 50 µL 100% H₂O₂) provides an excess of substrate, leading to uncontrolled bubble formation.
H₂O₂ Concentration: 100% H₂O₂ is highly reactive. Even small volumes can trigger violent reactions with catalase-rich samples.
Recommended Modifications
Reduce H₂O₂ Concentration:Dilute 100% H₂O₂ to 3–10% (commonly used in similar experiments) to slow the reaction.
Example: For 3% H₂O₂, mix 1.5 µL of 100% H₂O₂ with 48.5 µL distilled water.
Adjust Volumes:Decrease H₂O₂ volume while maintaining the sample volume (e.g., 15 µL liver tissue + 10–20 µL diluted H₂O₂).
Alternatively, reduce liver tissue volume (e.g., 5 µL) to limit catalase quantity.
Experimental Controls:Include a negative control (e.g., boiled liver tissue) to confirm catalase-dependent bubbling.
Add a surfactant (e.g., 1% Tween-20) to stabilize bubbles for better observation if needed.
I hope the above suggestions help you！
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