BD ELISPOT Reagents

BD Biosciences high-quality BD™ ELISPOT reagents offer the flexibility to choose among several formats: ready-to-use kits with pre-coated BD ELISPOT plates, coat-it-yourself sets, and matched antibody pairs. BD ELISPOT reagents coupled with a high-throughput ELISPOT plate reader provide objective and rapid analysis of cytokines at the single-cell level.


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..

BD ELISPOT Assay Procedure

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.


For Research Use Only. Not for use in diagnostic or therapeutic procedures.