BB2 Neptune Block Blocking Buffer

Neptune Block Blocking Buffer (BB2) is a non-mammalian blocking buffer designed primarily for antigen-down ELISA formats as well as sandwich assays with high background problems. BB2 is particularly useful when working with human and other mammalian serum samples as it works to reduce interactions between sample and blocking molecules.

Neptune Block Blocking Buffer is a mixture of small molecular proteins and molecular stabilizers capable of blocking the nonspecific binding sites of the adsorbed proteins, as well as the unoccupied regions of the plate that are not accessible to larger mammalian blocking reagents. When used in antigen-down ELISA formats, BB2's low molecular-weight proteins reduce the possibility of false-positives generated from endogenous antibodies in the sample reacting with plate-blocking proteins.

Neptune Block Blocking Buffer contains an antimicrobial agent for room temperature blocking. BB2 is formulated at 1X and is ready to use. It is stable for 18 months when stored at 2°-8°C or 1 month when stored at room temperature.

What does a block buffer do to the coated plate?
All of ICT's unique blocking buffers contain proprietary additives that stabilize the antigenic and functional regions of the coated protein by maintaining a level of hydration during the drying and storage process. These buffers also contain inert additives that block any "sticky" nonspecific binding regions of the adsorbed protein and any uncoated regions of the polystyrene plate surface. This reduces nonspecific binding of the conjugate and random deposits of sample proteins, reducing background noise and increasing the specific signal. ICT's proprietary block buffers also include an antimicrobial agent to prevent bacterial growth during the blocking process, which allows blocking to be done at room temperature, and to extend the shelf-life of the plates. Plates can be blocked and dried at room temperature and stored for 1 year or longer at 2°-8°C, depending on the coated protein.

When should BB2 be used?
BB2 is used as a high background blocker for primarily antigen-down assays as well as sandwich ELISA formats that require high level blocking. ICT offers a trial pack containing 100 mL each of three different blocking buffers for an economical and fast method of selecting the best blocking buffer for a particular assay (BBOP, catalog #957).

What type of ELISA plates will BB2 work on?
BB2 works on all types of polystyrene ELISA plates. For antigen-down ELISAs, ICT recommends Immulon® II High Binding plates, available from ICT (catalog #227); for antibody sandwich ELISAs, ICT recommends Corning Costar® 96-Well EIA/RIA Stripwell™ Plates (ICT catalog #25).

How are plates dried?
Plates coated and blocked with ICT's reagents should always be dried at room temperature. Blocked plates should first be aspirated to remove Block Buffer. To dry, place them in a fume hood, loosely covered with foil. Turn the fume hood on and dry overnight. An alternate method is to dry the plates in a vacuum chamber without heat for 4 hours. Plates should never be stacked on one another while drying. It is recommended that lyophilization should not be used as a drying method.

How should dried plates be stored?
Dried plates should be placed in a foil pouch (ICT catalog # 6288) with a 5 gram desiccant packet (ICT catalog # 6289) and sealed. The plates can then be stored at 2°-8°C for up to one year.