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Elispot

MultiScreen® HTS Filter Plates for Elispot

Optimized Filter Plates Measure Immune Response on a Single Cell Level

MultiScreenHTS filter plates are available with validated Immobilon®-P (PVDF) membrane for superior results. Plates with HA (mixed cellulose esters) are also available. Both provide membranes with dense uniform pore structure to promote antibody binding and increase sensitivity for better, sharper spot definition.

Image of a typical ELISPOT well
Image of a typical ELISPOT well
The spots are clear, focused and easy to distinguish. Photo courtesy of Zell Net Consulting, Inc.

Optimized Plate Design

Improved Spot Recovery and Spot Definition

Elispot assays are used for a range of applications to monitor immune responses and immunological response patterns, and can also be used to identify antigenic peptides and create vaccination strategies. Validated Elispot assays are highly reliable, reproducible and predictive. They can successfully identify antigenspecific T or B cells on a single cell level with results that closely mirror in vivo conditions and eliminate the need for long-term in vitro cell cultures or the addition of exogenous cytokines.

New MultiScreenHTS filter plates are specifically optimized for 96-well Elispot assays and are built to new specifications for improved performance.

Protocol

elispot protocol

  1. Coat membrane with antibodies. Add immune cells and incubate
  2. Responding cells produce cytokines. The cytokine of interest is then bound by the antibody
  3. Wash to remove cells. Add biotinylated antibodies which bind to the cytokine-antibody complex
  4. Add avidin-enzyme conjugate
  5. Add enzyme substrate and each responding cell will result in one spot

Reliable Assay Results

Reliable Assay Results

These images represent the number of cells secreting IFN-. in response to PHA-L stimulation of Human Peripheral Blood Mononuclearcytes (HPBMC). The wells were seeded with 50,000 cells and developed using BCIP/NBTplus substrate. The wells were imaged with the Zeiss KS Elispot imaging system. Typical MultiScreenHTS-IP filter plate variability expressed by %CV* is less than 10%. * %CV = (SD/mean)*100

Reliable Assay Results

MultiscreenHTS filter plates are designed for high performance Elispot assays. The plate design is optimized for improved membrane flatness to enhance in-well imaging. The plates are provided sterile and are constructed of low protein binding plastic. Each well is isolated and individually sealed to eliminate the occurance of cross talk. The plates are automation compatible and are in full compliance with ANSI/SBS 2004 standards. The plate design also features a removable underdrain for access to the membrane. Consistent results are seen within the plate and in plate-to-plate comparisons

출처: http://www.millipore.com