Intercept (TBS) Blocking Buffer

Intercept (PBS) Blocking Buffer

Specifications (Intercept Blocking Buffer)

• Intercept Blocking Buffer is a ready-to-use formulation. For optimal performance, do not dilute.
• Shelf Life: See expiration date on bottle.
• Storage: 4 °C
• Suitable for a variety of applications, including: quantitative Western blots, chemiluminescent Western blots, In‑Cell Western™ Assay, In-Gel Western, protein array, and glycoprotein detection.

Product Description (Intercept Blocking Buffer)

Blocking buffers enhance the quality of Western blots by reducing background interference, increasing the signal-to-noise ratio, and promoting specific binding of the primary antibody while minimizing non-specific interactions. Intercept Blocking Buffer is available in ready-to-use formulations in tris-buffered saline or phosphate-buffered saline to provide optimal blocking conditions for antibodies requiring TBS-based or PBS-based buffer systems.

The blocker does not contain mammalian proteins.

The TBS blocking buffer is ideal for use with antibodies against phosphorylated protein targets.

Materials Required for Blocking Membrane

• Blotted PVDF or nitrocellulose membrane

• Western blot incubation box

• Orbital shaker for incubation at room temperature

Guidelines

Use the same buffer system, PBS or TBS, for the entire Western blot process, including in wash solution.

Intercept T20 Antibody Diluents contain Tween® 20 and are available in TBS and PBS formulations, so you can use the appropriate buffer for your antibody diluent, without having to spend time measuring and adding Tween 20. When using a PVDF membrane, your secondary antibody diluent must contain a final concentration of 0.01 – 0.02% SDS.

Protocol

For a detailed Western blot protocol, see the Near-Infrared (NIR) Western Blot Detection Protocol (licorbio.com/wbanalysis). For troubleshooting ideas, see Good Westerns Gone Bad: Tips to Make Your NIR Western Blot Great (licorbio.com/GWGBIR).

Blocking Membrane

1. Place the PVDF or nitrocellulose membrane into the Western Blot Incubation Box and add undiluted Intercept Blocking Buffer to the membrane. Be sure to use sufficient blocking buffer to cover the membrane (a minimum of 0.4 mL/cm² is suggested).

   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.

2. Incubate for 1 hour at room temperature on an orbital shaker.

3. Proceed with Western blot detection protocol.

Related Products

See licorbio.com/intercept for ordering information.

Blocking Buffer

• Intercept (PBS) Protein-Free Blocking Buffer
• Intercept (TBS) Protein-Free Blocking Buffer

Antibody Diluent

• Intercept T20 (PBS) Antibody Diluent
• Intercept T20 (TBS) Antibody Diluent
• Intercept T20 (PBS) Protein-Free Antibody Diluent
• Intercept T20 (TBS) Protein-Free Antibody Diluent