Blocking Solutions/Background Suppression

Many compounds have been added empirically to immunolabeling solutions for the purpose of minimizing background staining. Researchers can obtain these items separately and prepare blocking and incubation media to suit the needs of their own protocols.

Blocking Solutions

The ready-to-use Blocking Solutions and the incubation media additive BSA-c™ are tuned for optimum background prevention and signal-to-noise ratio. In-depth information can be found in the respective product data sheets. There are also a number of components that allow researchers to formulate blocking, incubation and wash solutions according to the needs of their own experiments.

The signal-to-noise ratio determines the quality of any detection experiment. This ratio is in principle determined by the characteristics of specimen and detection reagents. With adequate blocking and incubation media, background reactions are minimized whereas specific reactions are not hampered.

Blocking Solutions effectively block 'sticky' surfaces by multi-point hydrophobic and charge-based interactions. The Blocking Solutions are specifically designed to meet the characteristics of each type of secondary conjugate.

Blocking Solutions are used to prevent immunoreagents from binding non-specifically to specimens with "sticky" surface properties.

Procedures to eliminate background comprise three main steps:

• To suppress residual aldehyde activity

• To saturate multipoint hydrophobic moieties and positive charges with high molecular weight compounds such as those present in the AURION Blocking Solutions

• To reduce aspecific binding of immunoreagents caused by hydrophilic interaction with competing molecules in the incubation and washing solution. AURION BSA-c™ is a particularly effective reagent for this purpose.

These steps should be balanced for optimum results. 

Specificity

Blocking Solutions are available in the following specificities:
serum-free: for use with Protein A and Protein G Gold conjugates.
with Normal Goat serum: for use with reagents based on secondary antibodies raised in Goat.
with Normal Rabbit serum: for use with reagents based on secondary antibodies raised in Rabbit.
with Normal Sheep serum: for use with reagents based on secondary antibodies raised in Sheep.
with Normal Donkey serum: for use with reagents based on secondary antibodies raised in Donkey.