BD ELISPOT Reagents

Features

• Direct analysis of frequency of cells producing cytokines and other effector molecules using enzyme-linked immunospot (ELISPOT) assay
• Greater sensitivity than ELISA and intracellular staining
• High-throughput analyses for direct ex vivo monitoring of T-cell function, including antigen-specific T cells

The ELISPOT assay is a powerful tool for detecting and analyzing individual cells that secrete a particular protein in vitro. Although originally developed for analyzing specific antibody-secreting cells, the assay has been adapted for measuring the frequencies of cells that produce and secrete a variety of other effector molecules such as cytokines.

BD ELISPOT kits, with pre-coated plates, sets, and pairs are available for human, non-human primate, and mouse research.

BD ELISPOT Reagent Pairs

Sufficient capture antibody and detection antibody for 5 plates.

BD ELISPOT Reagent Sets

Sufficient capture antibody, detection antibody, and streptavidin-horseradish peroxidase for 10 plates worth of assays (includes 10 uncoated plates).

BD ELISPOT Kits with Pre-Coated Plates

Designed for optimal spot quality, maximum time savings, and convenience, this kit includes all necessary reagents.

Kit contents:

• 2 antibody-coated plates
• Detection antibody
• Streptavidin-horseradish peroxidase
• AEC substrate/chromogen
• Wash concentrate
• PBS concentrate
• Assay diluent

BD ELISPOT Assay Procedures

Capture Antibody For sets and pairs: Coat microwells with anti-cytokine capture antibody. For Kits: Go to Step 3. Steps 1 and 2 are not necessary.
Blocking Block unoccupied sites with protein.
Add cells Incubate cells in well with Ag/stimulus, etc.
Wash Wash cells.
Detection Antibody Add biotinylated anti-cytokine detection antibody.
Enzyme-Streptavidin Add streptavidin-horseradish peroxidase.
Develop with substrate Add substrate and monitor formation of colored spots..

Features of BD ELISPOT assays

The BD ELISPOT assay is able to detect cytokine release at the single-cell level, allowing for direct determination of cytokine-producing cell frequencies. This assay has been found to be more sensitive than ELISA and intracellular staining. The sensitivity of the assay enables measurement of very low frequencies of cytokine-producing cells (eg, 1 in 300,000).

Recent developments in assay plate design and in high-throughput ELISPOT plate-reader instrumentation have significantly improved the usefulness of the ELISPOT method. Objective and rapid analyses of cytokine-producing cell numbers (spots) and relative amounts of cytokine produced per cell (spot size) are now possible.

BD ELISPOT assays enable high-sensitivity and high-throughput analyses for direct ex vivo monitoring of T-cell function, including antigen-specific T cells.

Quality antibody reagents result in crisp, clear results

Our BD ELISPOT reagents have been carefully screened to ensure the generation of crisp, clearly-defined immunospots. High-affinity BD Pharmingen™ NA/LE capture antibodies safeguard against nonspecific cellular activation. Detection antibodies have been carefully selected, along with pre-optimized preparations of streptavidin-horseradish peroxidase, substrate, and chromogen to produce optimal BD ELISPOT reagents.

Automated coating process ensures consistent results

For the kit format, BD Biosciences - Discovery Labware, the leader in biological coating and an ISO9001 facility, uses state-of-the-art automation to coat the BD ELISPOT plates to guarantee high-quality assay results with minimal well-to-well and plate-to-plate variation. The plates are coated with no azide/low endotoxin capture antibodies, are UV processed, and are tested and found negative for bacteria and fungi.