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Blocking Buffers

T293 blocking buffers

Figure 1. pAkt and ß-Tubulin antibody specificity depends on blocker selection. In 5% BSA, the pAkt antibody shows extra bands (green); in I-Block™ (Applied Biosystems), ß-Tubulin shows extra bands. In Odyssey Blocker (PBS), antibody specificity is increased.

LI-COR offers several blocking buffers for use with the Odyssey® and Aerius Systems. All are optimized for use with IRDye® products and other near-infrared fluorophores and provide excellent performance for quantitative Western blots and other immunoassays.

Blocker choice is important for immunoassay success. Blocking buffers enhance sensitivity by reducing background interference, increasing signal-to-noise ratio, and promoting specific binding of the primary antibody while minimizing non-specific interactions.

  • Insufficient blocking yields high background and reduced signal-to-noise ratios.
  • Excessive blocking may cause loss of blotted proteins1 or mask antibody:antigen interactions.
  • Detection reagents may cross-react with certain blocking buffers.
  • No single blocking buffer selection is suitable for ALL antigen-antibody pairs. Blocker choice may affect antibody specificity and non-specific binding2.
  • Empirical testing is critical! The Blocking Buffer Optimization Kit provides smaller quantities of three LI-COR blockers, to help you identify the best choice for your antigen.
  1. J Immunol Methods 1989, 122 (1), 129-35
  2. Proteomics 2008, 8, 2379–2383 [Figure 1]

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