Short Decsription
The purpose of ELISPOT analysis is to count cytokine producing cells in a single cell suspension. The advantage of this method is that it requires minimal in vitro manipulations, making the analysis of cytokine production as close to in vivo conditions as possible, and in a highly specific manner.
Description
The Elispot test is an ideal tool for TH1/TH2 response, vaccine development, viral infection monitoring and treatment, oncology, infectious diseases, autoimmune diseases and transplantation. Elispot detection is based on sandwich immunoenzyme technology. Cytokines or soluble molecules secreted by cells are captured by coating antibodies to avoid diffusion in the supernatant, protease degradation or binding to soluble membrane receptors. After the cells are removed, the captured cytokines are displayed by the tracer antibody and the appropriate conjugate.
Applications
ELISpot (ES)
Comment
Small volumes of IL-6 elispot set vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.
Molecular Mechanism of Action
After cell stimulation, locally produced cytokines are captured by IFNg and IL-4 specific monoclonal antibodies. After cell lysis, trapped cytokine molecules are revealed by a secondary anti-IFNg FITC conjugated antibody and a biotinylated anti-IL-4 antibody. Those are in turn recognised by anti-FITC HRP and streptavidin-AP conjugates. PVDF-bottomed-well plates are then incubated first with AEC substrate buffer, washed and subsequently incubated with BCIP/NBT. Coloured red/brownish spots indicate IFNg production while IL-4 is revealed by blue/purple spots.
Target
IL-6
Reactivity
Human
Detection Method
Sandwich
Method Type
Sandwich
Sample Type
cell suspension
Specificity
Recognizes natural human IL-6
Cross-Reactivity
No cross reactivity with other human cytokines. No interference with soluble receptor gp80.

For Research Use Only | Not For Clinical Use

Online Inquiry
Related Products
Copyright © 2024 Creative Biolabs. All Rights Reserved.