A recommended practice to reduce background staining in IHC is to **add a wash step with a mild detergent (like Tween-20) in the buffer**. Washing steps are critical throughout the IHC protocol to remove unbound antibodies and detection reagents that could contribute to non-specific background. Using a buffer, such as phosphate-buffered saline (PBS) or Tris-buffered saline (TBS), helps dilute and rinse away loosely bound molecules. Adding a small concentration (typically 0.05% - 0.1%) of a mild non-ionic detergent like Tween-20 or Triton X-100 to the wash buffer significantly improves its effectiveness. The detergent reduces surface tension, helps penetrate tissue folds, and disrupts hydrophobic and ionic interactions that can cause reagents to stick non-specifically to the glass slide or tissue. Thorough and vigorous washing after each incubation step (primary antibody, secondary antibody, enzyme complex) is one of the simplest and most effective ways to achieve a clean background, making the specific signal stand out clearly.

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Zambia

• Joseph Soko – 101 pts

• Visual Pathway Lesions
• Positive and Negative Symptoms
• Hallucinations and Delusions
• Antipsychotic Medications
• Emergency: Environmental Emergencies