Intercept Blocking Buffers

Intercept® Blocking Buffer Answer Portal

For example, "reconstitute secondary antibody".

Safety Data Sheets (SDS) are available on the SDS Search Page.

Intercept Blocking Buffers are formulated to provide highly efficient blocking and low background variability. 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.

Intercept Protein-Based Blocking Buffers (TBS and PBS)

  • Description: non-mammalian, protein-based

  • Uses: 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.

  • Form: 1X Concentration

  • Example Supported Application: Quantitative Western Blot, Chemiluminescent Western Blot, In‑Cell Western™ Assay, In-Gel Western

Intercept Protein-Free Blocking Buffers (TBS and PBS)

  • Description: Proprietary protein-free blocking buffer

  • Uses: 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.

  • Form: 1X Concentration

  • Example Supported Application: Quantitative Western Blot, Chemiluminescent Western Blot, In‑Cell Western Assay, In-Gel Western

Tips

  • For best results, do not dilute.

    Shake well before each use.

  • Do not use detergents like Tween® 20 or SDS during the blocking step, as this may generate a background signal. Detergents should only be added when diluting the primary and secondary antibodies.

Frequently Asked Questions

Which blocking buffer do I use?

Blocking buffer choice is important for immunoassay success. Keep these considerations in mind when choosing a blocking buffer.

What should I do if my blocking buffer got frozen or it sat on the bench for too long?

If you have concerns, it is safest to replace the blocking buffer.

If the blocking buffer was frozen, you can try thawing in a fridge. If the blocking buffer has been outside the recommended 4 °C for too long, it may be unusable.

How can I minimize background?

Avoid non-specific background staining on PVDF membranes: If you are making your own antibody diluent, add SDS (final concentration 0.01 – 0.02%) and Tween® 20 (final concentration 0.2%) to the diluent. If using Intercept® T20 Antibody Diluent, only add SDS (final concentration 0.01 – 0.02%).