Blocking Buffers


LI-COR offers several blocking buffers for use with the Odyssey® Imaging 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.

Choose the Right Blocking Buffer for Your Application

Blocking buffer choice is important for immunoassay success. Blockers 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.
  • The Blocking Buffer Optimization Kit provides smaller quantities of three LI-COR blocking buffers, which can help empirically identify the best choice for your antigen.
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), β-Tubulin shows extra bands. In Odyssey Blocker (PBS), antibody specificity is increased.

LI-COR Blocking Buffers can be used in many applications, including:

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Blocking Buffers for Western Blotting

Sign up to review the science, best practices, and mechanics of the blocking buffer step for either chemiluminescent or near-infrared fluorescent Western blots at Lambda U online learning portal.

These online, on-demand resources can help ensure that you are getting the very best Western blot data.

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  1. DenHollander, N., and Befus, D. (1989). Journal of Immunological Methods, 122(1), pp 129-135.
  2. Ambroz, K.L., Zhang, Y., Schutz-Geschwender, A., and Olive, D.M. (2008). Protemoics, 8(12), pp 2379-2383. DOI: 10.1002/pmic.200700676.
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