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ELISpot Assays

 Measure cellular response quantitatively – with single-cell precision

ELISpot assays are used in a range of applications to monitor immunological response patterns, and their utility for the identification of antigenic peptides has proven useful for the development of immunotherapeutics and vaccine strategies. Validated ELISpot assays are highly reliable, reproducible and predictive. They can successfully identify antigen-specific T or B cells at the single cell level with results that closely simulate in vivo conditions, and can eliminate the need for long-term in vitro cell cultures.

ELISpot combines plate-based Enzyme Linked Immunosorbent Assays (ELISAs) with membrane-based Western blotting technologies for assessing immune cell function based on secreted analytes. T and B lymphocyte activation and related responses are key to clinical outcome in cancer and in a variety of other human diseases, but simultaneous measurement of key secreted factors can present a technical challenge. The ELISpot technique enables multi-target, quantitative assessment of effector functions at the single cell level, with exceptional sensitivity and resolution.

Membrane characteristics and application validation are key for this precise immunodetection technique.  We are proud to be the leading supplier of validated ELISpot assays that incorporate unsurpassed Millipore membranes to the life science and biotherapeutic development communities.

 

 The ELISpot Assay Workflow

The ELISpot Assay Workflow

  1. Coat membrane with capture antibody. Add immune cells and stimuli.
  2. Responding cells produce cytokines. The cytokine of interest binds to the capture Ab.
  3. Wash away cells. Add a second cytokine-specific biotinylated Ab, which binds the cytokine-Ab complex.
  4. Add enzyme substrate and develop. Each spot within a well represents the cytokine signature of a single cell.

The ELISpot technique can be used to detect diverse cellular responses that include

  • cytolytic activity
  • secretion of multiple cytokines and other bioactive molecules
  • proliferation

 

Select ELISpot Applications and Publications

Disease Recent ELISpot related reviews
Alpha-1 antitrypsin deficiency (AAT) Flotte, T. and Mueller, C. (2011) Hum Mol Genet R1:R87-R92.
BK Virus /nephropathy Sawinski, D. and Goral, S. (2015) Nephrol Dial transplant 30: 209-217.
Diabetes Culina, S. et al. (2013) Eur J Endocrinol 168:R19-R31.
Genetic disease/ AAV therapy Mingozzi, F. and High, K. (2013) Blood 122:23-36.
Hepatitis Krain, L. et al. (2014) Clin Microbiol Rev 27:139-165.
HIV Van de Perre, P. et al. (2012) Science Translational Medicine 4:143sr3.
Lung cancer Kao, CJ et al. (2014) Cancer Immunol Res 2:581-589.
Lymphoma Brody, J. et al. (2011) J Clin Oncol 29:1864-1875.
Malaria McCall, M. and Sauerwein, R. (2010) J. Leukoc Biol 88:1131-1143.
Multiple Myeloma Fichtner, S. et al. (2014) Clin Cancer Res 21:1712-1721.
Neuroinflammation Figuera-Losada, M. et al. (2014) J Biomol Screen 19:17-31.
Prostate Cancer Singh, B. and Gulley, J. (2014) Therapeutic Advances in Vaccines 2:137-148.
Salmonella Bhuiyan, S. et al. (2014) Am J Trop Med Hyg 90:1024-1030.
Tick-borne encephalitis (TBE) Schwaiger, J. et al. (2014) J Virol 88:7828-7842.
Tuberculosis (TB) Hawn, T et al. (2014) Microbiol Mol Biol Rev 78:650-671.
Viral myocarditis Yang, L. et al. (2014) Clin Vaccine Immunol 21:312-320.

 

 Ordering Information

MultiScreen® HTS 96-well Plates*
 

Description Plate material/
color
Qty/Pk Sterile Cat. No.
MultiScreen® HTS-HA plates with MCE membrane Styrene/clear
10 Yes MSHAS4510
MultiScreen® HTS-IP plates with Immobilon®-P membrane Acrylic/clear
Acrylic/clear
Acrylic/white
Acrylic/white
10
50
10
50
Yes
No
Yes No
MSIPS4510
MSIPN4550
MSIPS4W10 MSIPN4W50
MultiScreen® plates with Immobilon®-P membrane without underdrain   Acrylic/white No underdrain 10 Yes MAIPSWU10
MultiScreen® HTS plates with Immobilon®-P-FL membrane, with underdrain Acrylic/white 10 Yes S5EJ104I07

* Classic MultiScreen® plates are also available for ELISpot assays. Contact technical service for information on catalogue numbers MAHAS4510, MAIPS4510 and S2EM004M99.

 

8-well strip
 

Description Plate material/
color
Qty/Pk Sterile Cat. No.
MultiScreen® 8-well strip with Immobilon®-P membrane Acrylic/Clear 10 Yes M8IPS4510

 

Accessories
 

Description Qty/Pk Cat. No.
Plate sealing tape, opaque 100 MATAH0P00
MultiScreen® 8-Well Strip Support Frame 10 M8IPFRAME