Improve the performance of your assay.
ImmunoChemistry Technologies (ICT) offers a comprehensive line of high-quality buffers, diluents, and solutions for the preparation and execution of ELISA tests. From start to finish, these reliable assay solutions control variables and address complex issues including matrix effects, non-specific binding, and signal-to-noise ratio. These reagents have been specifically optimized for a 96-well microtiter plate system, and can be applied in other assay techniques as well.
ICT offers reliable ELISA solutions:
The first step in making a reliable ELISA is proper coating of the antibody or antigen onto the plate. ICT has developed 2 coating buffers based on the type of ELISA being made. ICTs 5x Universal Antibody Plate Coating Buffer (CB1) is a unique buffer used to coat antibodies onto polystyrene microtiter ELISA plates, and can be used for antibody-sandwich ELISAs as well as antigen-down ELISAs. ICTs 5x Antigen Coating Buffer (CB2) is designed for antigen-down ELISAs. Both coating buffers stabilize coated proteins by maintaining their tertiary three-dimensional structure, allowing for greater binding reactivity with the detection molecule, thereby enhancing the specific signal.
Once the antibody or protein antigens have been properly adsorbed onto the plate (we suggest using ICT’s Plate Coating Buffer, CB1), the next critical step in creating a reliable assay is the blocking of the plate. Blocking is done to protect the coated protein from harsh external conditions while masking any uncoated regions on the plate. As the blocking process has a profound effect on both the specific and nonspecific signals generated in the assay, the proper selection of block buffer is a key element in the preparation of a functional ELISA. The proper block buffer can increase sensitivity, reduce nonspecific binding, and lengthen the shelf-life of the coated plate. Because ELISAs may be configured in several ways, each with unique blocking requirements, ICT has created 4 proprietary block buffer formulations for use with sandwich and antigen-down ELISAs. They are:
BB1 General Low-Level Blocker with BSA.
BB2 Neptune Block with Nonmammalian (fish) proteins, for extra blocking strength.
BB3 SynBlock, based on small synthetic non-protein blocking molecules, for ELISAs that require extra blocking strength.
BB4 Phosph-Free Blocker, does not contain any phosphates nor any animal proteins. It is ideal for ELISAs using alkaline phosphatase detection systems, and ELISA with super sensitivity requirements.
If you would like to test our first three blockers in your assay system, just buy the optimization pack: it includes 1 100mL bottle of each BB1, BB2, and BB3.
ICT has formulated a group of unique stabilizing diluents designed to prolong the shelf-life of HRP-conjugated proteins and enhance their utility in ELISA applications. These conjugate stabilizer diluents can be used to reconstitute lyophilized conjugates, and to dilute concentrated conjugates into the useful range of the assay. HRP-conjugated antibodies are often unstable and can lose a significant level of activity within a short time after rehydration and dilution unless properly stored. This loss of activity is based on two factors: denaturization of the HRP-IgG protein complex; and destabilization of the catalytic region of the HRP. ICT’s proprietary formulations preserve the integrity of the antibody binding regions while maintaining the enzymatic functionality of the HRP enzyme. ICT has created 6 proprietary conjugate diluent formulations for use with sandwich and antigen-down ELISAs, which are all available at 1x and 5x concentrations:
CS1 Monoclonal/Polyclonal Conjugate Diluent & Stabilizer.
CS2 Antigen-Down Conjugate Diluent & Stabilizer.
CS3 Polyclonal/Polyclonal Conjugate Diluent & Stabilizer.
CS4 Monoclonal/Goat- Polyclonal Conjugate Diluent & Stabilizer.
CS5 Monoclonal /Rabbit- Polyclonal Conjugate Diluent & Stabilizer.
CS6 Conjugate Stock Stabilizing Reagent.
ICT’s assay diluents are proprietary buffer formulations designed to equalize any differences between the sample matrix (serum, plasma, urine, cell culture fluid) and the diluent used to generate the standard curve. These diluents also reduce nonspecific interactions between the sample matrix proteins and the plate surface which translates into lower background noise. Assay diluents are pipetted directly onto the plate just prior to adding the samples, and may be used with neat or diluted samples, depending on the characteristics of the sample matrix and the target range of the assay. Since different sample matrices have unique characteristics when run in an ELISA, they must be matched with specific assay diluents. Use of an assay diluent reduces the effects of the sample matrix and variation among samples, without pre-dilution of the samples. ICT has created 4 proprietary assay diluent formulations for use with sandwich and antigen-down ELISAs:
AD1 General Assay Diluent for Serum & Plasma Samples.
AD2 IgM-Reducing (Positive Interference) Assay Diluent for Serum & Plasma Samples.
AD3 Neptune Assay Diluent for Serum & Plasma samples.
AD4 Antigen-Down Assay Diluent for Serum and Plasma samples.
If you would like to test all four in your assay, buy the optimization pack: it includes 1 100mL bottle of each AD.
ICT’s unique serum sample diluents are specifically formulated for dilution of animal and bird serum samples in antigen-down ELISA formats. Sample diluents are utilized to dilute serum or antigen samples into the functional range of the assay. Due to the finite binding capacity of the coated ELISA plate surface, certain high samples may overload this capacity, thus requiring dilution prior to testing in the assay. Sample diluents can also reduce background noise associated with nonspecific bridging of signal-generating conjugates to the plate surface. Hyperimmunized serum samples contain nonspecific IgG antibody that may interfere with the titration signal. Proprietary additives have been incorporated into these diluents that minimize nonspecific binding interactions of the nonspecific IgG in the samples, thus decreasing background noise. These sample diluents can be used to titrate rabbit, mouse, goat, and chicken serum samples, along with monoclonal cell culture supernatants. Antigen samples may also be diluted with sample diluents for evaluation in sandwich ELISA formats. ICT has created 3 proprietary sample diluent formulations for use with antigen-down and some sandwich ELISA applications:
SD1 General Sample Diluent for Serum, Ascites, and Cell Culture Supernatant Samples.
SD2 Plasma Sample Diluent for Antigen-Down testing of Plasma Samples.
SD3 Neptune Sample Diluent.
If you would like to test all three in your assay, buy the optimization pack: it includes 1 100mL bottle of each SD.
ICT’s Universal ELISA Wash Buffer is a well-tested formulation of buffers, salts, and detergents designed to effectively remove excess material from microtiter plates without disrupting ELISA binding reactions. By maintaining the proper buffering environment, unbound components can be washed away without suppressing antigen-antibody binding interactions, thereby reducing nonspecific background noise and increasing the specific signal. ICT’s Wash Buffer is compatible with all routinely used conjugate components such as HRP, AP, and avidin, among others.
ICT’s 10X Phosphate Buffered Saline is a well-tested formulation of buffers and salts designed to effectively balance the pH without disrupting ELISA binding reactions.
Also use it as a base to create your own ELISA buffers or for other applications in the lab like dialyzing proteins and running samples over HPLC and Protein G columns.