Conductivity Testing

CAPS buffers effectively in the alkaline range from about pH 9.7 to 11.1, with optimal buffering near pH 10.4.

Selecting the right blocking buffer is essential for reducing background noise and ensuring strong, specific antibody binding in your SDS-PAGE and western blot protocol. Which one you choose depends on your target protein, antibody type, and detection method.

• Non-Fat Powdered Milk: This is a cost-effective and widely used option for general Western blot applications. It works well with most primary and secondary antibodies but can interfere with phospho-specific antibodies due to its protein content.
• Bovine Serum Albumin: Ideal for phospho-specific and glycoprotein antibodies, as it lacks phosphoproteins that could cause background issues. It is also a good alternative when milk-based blocking buffers fail.
• Casein: A purified milk protein that provides effective blocking while minimizing cross-reactivity. This is good for fluorescent Western blots and when using certain monoclonal antibodies.
• Gelatin: A good option for reducing background in fluorescent and chemiluminescent detection. It works well with antibodies that show non-specific binding in milk or BSA.

Low concentrations of Tween-20 (0.05–0.1%) in PBS or TBS help remove unbound antibodies, reducing background and improving signal clarity.

Buffers are used in HPLC to mitigate the influence of ionization of acidic or basic functional groups of analytes being separated. The buffer reduces fluctuations of pH, which would consequently cause unwanted influence on separation of the analytes.