Background Is Controlled by Three Independent Steps
- Low Molecular Weight Block (before protein block)
Purpose: to inactivate residual fixative e.g. aldehydes, using:
– amino acids such as glycin or lysin, or
– aldehyde inactivating compounds such as NaBH4 and NH2OH
- High Molecular Weight Protein Block (before immunolabeling)
Purpose: to prevent stickiness to hydrophobic areas and domains with excessive positive charges based on multiple point interactions (high affinity protein binding capacity), using:
– normal serum
- Incubation and Wash solution (during immunolabelling)
Purpose: to eliminate aspecific binding of immunoconjugates based on hydrophilic interactions (“oligo”point interactions) by competition, using:
acetylated albumin (AURION BSA-c™) in the incubation buffer
Please refer to our Newsletter 1 and Newsflyer 1 for more information on the subject of background.
You can find incubation protocols in the section on this website.
Abandoning BSA-c™ molecules compete with antibodies and secondary immuno reagents for aspecific binding to positively charged specimen compounds, thus minimizing background.