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Human IL-10 ELISPOT Kit (with un-coated plates) is a ELISPOT kit for the sensitive detection of Human IL-10 production in a single cell suspension.

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Key facts

Detection method
Colorimetric
Sample types
Suspension cells
Assay type
Sandwich (qualitative)
Reactive species
Human, Monkey

Reactivity data

Application
ELISPOT
Reactivity
Reacts
Dilution info
-
Notes

-

Target data

Function

Major immune regulatory cytokine that acts on many cells of the immune system where it has profound anti-inflammatory functions, limiting excessive tissue disruption caused by inflammation. Mechanistically, IL10 binds to its heterotetrameric receptor comprising IL10RA and IL10RB leading to JAK1 and STAT2-mediated phosphorylation of STAT3 (PubMed:16982608). In turn, STAT3 translocates to the nucleus where it drives expression of anti-inflammatory mediators (PubMed:18025162). Targets antigen-presenting cells (APCs) such as macrophages and monocytes and inhibits their release of pro-inflammatory cytokines including granulocyte-macrophage colony-stimulating factor /GM-CSF, granulocyte colony-stimulating factor/G-CSF, IL-1 alpha, IL-1 beta, IL-6, IL-8 and TNF-alpha (PubMed:11564774, PubMed:1940799, PubMed:7512027). Interferes also with antigen presentation by reducing the expression of MHC-class II and co-stimulatory molecules, thereby inhibiting their ability to induce T cell activation (PubMed:8144879). In addition, controls the inflammatory response of macrophages by reprogramming essential metabolic pathways including mTOR signaling (By similarity).

Alternative names

What's included?

5 x 96 Tests
Components
96 PVDF-bottomed-well plates.
5 x 1 Unit
Biotinylated detection antibody, clone B-T10
1 x 1 Vial
Bovine Serum Albumin
1 x 2 g
Human IL-10 Capture antibody
1 x 500 µL
Ready-to-use BCIP/NBT substrate buffer
2 x 25 mL
Streptavidin - Alkaline Phosphatase conjugated
1 x 50 µL

Recommended products

Human IL-10 ELISPOT Kit (with un-coated plates) is a ELISPOT kit for the sensitive detection of Human IL-10 production in a single cell suspension.

Key facts

Detection method
Colorimetric
Sample types
Suspension cells
Assay type
Sandwich (qualitative)
Reactive species
Human, Monkey
Assay Platform
Microplate

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C

Notes

The Human IL-10 ELISPOT Kit (with un-coated plates) is designed to enumerate cytokine producing cells in a single cell suspension. This method has the advantage of requiring a minimum of in-vitro manipulations allowing cytokine production analysis as close as possible to in-vivo conditions in a highly specific way. This technique is designed to determine the frequency of cytokine producing cells under a given stimulation, and the follow-up of such frequency during a treatment and/or a pathological state. Elispot assay constitutes an ideal tool in the TH1 / TH2 response, vaccine development, viral infection monitoring and treatment, cancerology, infectious diseases, autoimmune diseases and transplantation.

This ELISPOT assay is based on sandwich immuno-enzyme technology. Cell secreted cytokines or soluble molecules are captured by coated antibodies avoiding diffusion in supernatant, protease degradation or binding on soluble membrane receptors. After cell removal, the captured cytokines are revealed by tracer antibodies and appropriate conjugates.

Principal:

After cell stimulation, locally produced cytokines are captured by a specific monoclonal antibody. After cell lysis, trapped cytokine molecules are revealed by a secondary biotinylated detection antibody, which is in turn recognised by streptavidin conjugated to alkaline phosphatase. PVDF-bottomed-well plates are then incubated with BCIP/NBT substrate. Colored "purple" spots indicate cytokine production by individual cells.

Recognizes natural human IL-10

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