Custom Premix Selecting "Custom Premix" option means that all of the beads you have chosen will be premixed in manufacturing before the kit is sent to you.
A ready-to-use blocking reagent for Western blotting and immunoassays. Contains salmon plasma proteins that do not interact with mammalian antibodies, thus offering a reduced background.
Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.
Revision
08-August-2022 JSW
Description
A ready-to-use blocking reagent containing non-mammalian proteins that do not interact with mammalian antibodies.
Form
Liquid
Formulation
In PBS, 0.09% NaN₃, pH 7.4.
Recommended reaction conditions
Use at 1X concentration or further dilution depending on specific use.
BioAssay results using Aquatic Block™ Reagent as a blocking reagent
Comparison of Aquatic Block™ Reagent to standard blockers: 3% BSA, 0.2% Non Fat Dry Milk, and 10% Fetal Bovine Serum.
All assays were performed following standard protocols, except the blocking reagent was added prior to the addition of the antigen and/or antibody.
All measurements reported as average Absorbance. The blank well was subtracted from the actual readings.
Table 1: Sample Wells
Medium Binding: All the blocking reagents exhibit excellent blocking ability on the surface-BSA being the best performer by a slight margin. Note: blocking mechanism for this surface is hydrophobic.
High Binding: All the blocking reagents exhibit excellent blocking ability on this surface-FBS being the best performer by a very small margin. BSA, the only single protein type blocker, exhibited the least effective blocking (still excellent) as compared to the other blockers. Since a high binding surface is a combination of hydrophobic and ionic binding mechanisms, a blocker comprised of several proteins typically performs better.
Glutaraldehyde Activated Amine (Polystyrene): BSA is a poor blocker for this surface. Typically a covalent surface such as this one (combination of Hydrophobic, ionic and covalent binding mechanisms) requires a multicomponent blocking reagent- such as a normal serum. The best blocker for this surface was fish serum, followed by fetal bovine serum and non-fat dry milk.
N-oxysuccinimide: This covalent surface is comprised of NOS groups and hydrophobic spacer arms. FBS was the best performer, fish serum and NFDM also performed adequately.
None of these blocking reagents can be promoted as universal blockers, since they are surface dependent.
Table 2: Ranking per Surface Type: Best to Worst
Note: There are only very minor differences in many cases between the blockers and one can not call one better than the other. It is obvious that BSA is the poorest blocker, although it is highly promoted as a "superior" blocking reagent. Fish serum blocked all of the surfaces very well.
