Total IgG secreting B cells produced by 2x10E5 rhesus macaque cells/well.
5-plate red spots (plates NOT included)
Monkey IgG B cell ELISPOT Kit
INTENDED USE
The B cell ELISPOT (Enzyme-linked Immunospot) assay has been designed to identify and enumerate individual antibody secreting cells (ASC) in single cell suspensions of (most commonly) peripheral blood mononuclear cells (PBMC).
BRIEF DESCRIPTION OF B CELL ELISPOT ASSAY
A cell suspension with B cells are brought into the wells of the coated ELISPOT plate and incubated for a defined length of time at 37°C. Antibodies released by the B cells are captured by the coated Ig or antigen. After incubation, cells are washed away and areas in which secreted antibodies are bound are detected by the sequential addition of biotinylated anti-isotype specific antibodies and an enzyme conjugate or Ψ-labeled anti-biotin antibodies (GABA). The last step in the assay is the addition of AEC substrate (3-amino-9-ethylcarbazole) or a reagent allowing the precipitation of silver on Ψ revealing the sites of antibody secretion (footprints of individual ASC). These footprints (spots) represent either the total number of ASC (in Ig-coated wells) or antigen-specific ASC (in antigen-coated wells).
- Coating antibodies (to determine total number IgG secreting B cells)
- Biotinylated detector antibodies
- B cell stimulus
- Streptavidin-HRP conjugate
- AEC coloring system
- Blocking stock solution R
- Dilution buffer R
- Tween-20
- Detailed manual
Streamline laboratory procedures! Click here for ready-to-use reagents and buffers, from Medicago AB. Available in tablets and pouches!
