Nucleic Acid Binding, Loading & Release

Heparin Competition Assay

Complex stability against polyanions that mimic the extracellular matrix and serum.

Purpose

To assess the stability of nucleic acid–carrier complexes in the presence of polyanions (mimics the extracellular matrix or serum proteins).

Standard protocol

1
Prepare fully complexed nanoparticles (at the optimal N/P ratio).
2
Add increasing concentrations of heparin (e.g., 0.1–5 U/mL).
3
Incubate 15–30 min at 37 °C.
4
Run on agarose gel (as in the gel retardation assay).
5
Observe: if a band reappears, the nucleic acid was released due to competition.

Agarose gel of complexes challenged with increasing heparin concentrations; reappearing bands indicate displaced nucleic acid.

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