Rat IL-4 ELISPOT Kit w/o plates

Molecule:

IL-4

Species:

Rat

Cat. No.:

CKR034 pdf (datasheet)

Quantity/Size:

5 Plates

Price:

$1,330.00 BUY

Description:

Intended use
The cytokine ELISPOT assay is designed to enumerate cytokine secreting cells in single cell suspensions of lymphoid tissue, CNS tissue, bone marrow or preparations of peripheral blood mononuclear cells (PBMC). The assay has the advantage of detecting only activated/memory T-cells and the cytokine release can be detected at the single cell level, allowing direct determination of T-cell frequencies. The high sensitivity and easy performance, allowing a direct enumeration of peptide-reactive T-cells without prior in vitro expansion, makes the ELISPOT assay eminently well suited to monitor T-cell responses. The higher sensitivity if ELISPOT in comparison to that of ELISA or intracellular staining is due to the plate-bound antibodies directly capturing the cytokine released by the cell before it is diluted in the supernatant, trapped by high-affinity receptors or degraded by proteases. The sensitivity of the assay lends itself to measurement of very low frequencies of cytokine-secreting cells (1/300,000).

Brief description of the ELISPOT assay
Cells are incubated in the wells of the ELISPOT plate precoated with a high-affinity monoclonal antibody to which the cytokine, produced during incubation, will bind. Subsequently, cells are washed away. Areas in which the cytokine has been trapped are detected with a combination of biotinylated anti-cytokine detection antibodies and Streptavidinhorseradish peroxidase (Streptavidin-HRP). The last step in the assay is the addition of AEC (3-amino-9-ethylcarbazole) yielding a red zone (“spot”). This zone reveals the site of cytokine secretion.

Reagents Included

  • Coating antibody, lyophilized
  • Biotinylated detection antibody, lyophilized
  • Streptavidin-HRP conjugate, lyophilized
  • AEC stock solution
  • AEC Substrate buffer capsules
  • Blocking stock solution (10x)
  • Dilution buffer R (10x)
  • Tween-20

Data PDF:

CKR034