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Milk can leave a thick protein deposit, resulting in non-specific binding of the antibody to the entire blot (top panel). Bløk® reagent coats the blot with a thin chemical layer that does not bind antibodies (bottom panel), leading to less non-specific binding by the antibodies and lower background.
In Western blotting, blocking of unbound membrane sites is necessary to prevent non-specific binding of the antibodies, which leads to high backgrounds.
Traditional milk/protein-blockers can leave a thick layer of sticky proteins that:
Promotes non-specific interactions with antibodies, leading to an increased background.
Are not compatible with detection of protein phosphorylation due to the presence of phosphoproteins in milk.
Bløk® chemical-based, protein-free blocker decreases background caused by non-specific antibody binding without leaving a thick, sticky layer similar to milk.
Bløk® Noise Cancelling Reagents are designed especially for Western blotting applications. This family of protein-free, ready-to-use buffers has been optimized to reduce background levels when using chemiluminescence, fluorescent, or phosphotyrosine detection.