Before membrane bound antigens can be detected, unspecific binding sites for
antibodies on the transfer membrane have to be blocked. For many applications skim
milk powder is an economic alternative to bovine serum albumin (BSA). For
Western Blots where skim milk cannot be used, SERVA offers highly purified
bovine serum albumins.
Protein-containing blocking solutions, such as skim milk powder or BSA solutions, may not only block non-specific antibody binding sites, but also mask specific binding sites. BlueBlock PF, a protein-free, polymer-based blocking reagent, enables the blocking of non-specific binding sites in only 5 min without loss of sensitivity in colorimetric, chemiluminescence and fluorescence detection systems. The All-in-One protocol, the antibody incubation in the blocking solution, allows the procedure to be performed in only 1 - 2 hours.
After blotting of nucleic acids before hybridization, non-specific binding sites are blocked by Denhardt’s solution (cat. no. 39603).
In addition to a
broad range of substrates as powder, SERVA offers ready-to-use, highly
sensitive substrate solutions for easy and fast detection of proteins in
SERVALight HRP Chemiluminescence Kits are substrates for the detection of membrane bound antigens (Western Blot) or nucleic acid sequences (Southern and Northern Blot), labelled directly with Horseradish Peroxidase (HRP) or indirectly with HRP-conjugated antibodies/streptavidin. They have an excellent stability, extended signal duration and save money and precious antibodies due to high dilution of antibodies.
SERVAColor BCIP/NBT and TMB Blot Solution are non-toxic, highly sensitive substrate solutions for the colorimetric detection of alkaline phosphatase (AP) or horseradish peroxidase (HRP) in membrane assays.
The testing of new primary antibodies or optimal antibody concentrations requires the performance of several Western blots. This is not only expensive, but also wastes rare or expensive samples. By removing the antibodies ("stripping") after chemiluminescence detection and reusing them ("re-probing"), the use of several gels to examine different target structures can be avoided. BlueClear SB is a ready-to-use buffer for the efficient stripping of high-affinity antibodies from Western blot membranes. BlueClear SB Stripping Buffer removes primary and secondary antibodies cleanly and efficiently from Western blots without loss or damage of the immobilised antigen, allowing reliable re-hybridisation of the blots.