Anti-mouse IL-17A mAb (MT2270), biotin

Anti‑mouse IL‑17A mAb (MT2270), biotin

3521-14-250

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Content

Monoclonal antibody MT2270, biotinylated. Supplied at 0.5 mg/ml in PBS with 0.02% sodium azide.

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Product specifications

Intended use

This monoclonal antibody enables specific detection of mouse IL-17A in immunoassays such as ELISpot, FluoroSpot, and ELISA.

Serum/Plasma samples

Recommendation

MT2270 is recommended as detection mAb in ELISpot, FluoroSpot, and ELISA in combination with capture mAb IL17-I (product code 3521-3).

Product details

ProductAnti-mouse IL-17A mAb (MT2270), biotin
ApplicationELISpot, FluoroSpot, ELISA
AnalyteIL-17A
AntibodyMT2270
ConjugateBiotin
ClonalityMonoclonal
ImmunogenRecombinant mouse IL-17A
HostMouse
IsotypeIgG1
ReactivityMouse
SpecificityNative and recombinant mouse IL-17A
PurificationPurified from in vitro cultures by protein G affinity chromatography.
BiotinylationBiotinylated through reaction with a N-hydroxysuccinimide ester of biotin.
Concentration0.5 mg/ml
Supplied inPBS with 0.02% sodium azide. Sterile-filtered (0.2 µm).
ContentsMonoclonal antibody MT2270, biotinylated. Supplied at 0.5 mg/ml in PBS with 0.02% sodium azide.

Shipping and Storage

ShippingShipped at ambient temperature.
StorageStore product at 4-8°C or frozen at -20°C or below. Avoid repeated freezing/thawing.
Shelf lifeAt least 18 months from date of receipt.

Find out which analyte combinations we have evaluated in T cell FluoroSpot and which combinations are affected by capture effects or not. 

What is a capture effect?

When capture antibodies with different specificities are coated together, the capture of one cytokine may affect the secretion of other cytokines. This is usually more pronounced when studying T cell responses with polyclonal stimuli compared to antigen-specific responses. Capture effects are seldom a problem and can often be counteracted by the addition of an anti-CD28 antibody.

Depiction of capture effects observed in FluoroSpot

(1) IL-2 secreted by the activated T cell is captured by coated anti-IL-2 capture antibodies. (2) As a result, IL-2-stimulation of the T cell itself (autocrine stimulation) as well as nearby T cells (paracrine stimulation) is impaired, ultimately leading to (3) decreased IFN-γ secretion.

Co-stimulation with anti-CD28

Anti-CD28 mAb provides a co-stimulatory signal to antigen-specific responses by binding to CD28 on T cells. The addition of an anti-CD28 mAb to the cell culture enhances antigen-specific responses and can counteract capture effects. For example, the presence of IL-2 capture antibodies may result in reduced activation of T cells, as capturing of IL-2 decreases the amount of available IL-2 and thereby dampens the IL-2-mediated stimulation of T cells. The addition of anti-CD28 mAb restores IFN-γ responses (depicted in the below images). Further optimization may be necessary, depending on the cells and stimuli used. Too high concentrations of the anti-CD28 mAb may result in an elevation of non-specific cytokine secretion.

Overcoming capture effects using anti-CD28 anitbodies in FluoroSpot

(1) An anti-CD28 antibody can be added to provide a co-stimulatory signal that can restore (2) e.g. IFN-γ responses.

How to investigate capture effects

Our FluoroSpot Plus kits are evaluated for capture effects, and in studies of T cell responses we recommend co-stimulation with anti-CD28. With FluoroSpot Flex, it is possible to combine and build your own kit. For guidance look at our analyte combination table (above). Capture effects can be investigated by quantifying spot numbers in wells coated with single capture antibodies and compared to wells coated with a mixture of the capture antibodies. The compensatory effect of the anti-CD28 mAb may be assessed by comparing cells cultured with and without the anti-CD28 mAb.

Tutorial, Published December 28, 2023

Incubation strategies for success in ELISpot and FluoroSpot
We've made a handy table summarizing incubation times for different analytes to make your research planning for the ELISpot and FluoroSpot assay a little easier.
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IL-17A

Analyte description

Interleukin 17A (IL-17A) is a potent proinflammatory cytokine produced by activated Th17 (T helper 17) cells and certain cells belonging to the innate immune system. In mice, IL-17 has also been shown to be produced by activated CD8 T cells and γδ T cells. Th17 cells play an important role in autoimmune diseases and protection against bacteria and fungi. IL-17A acts on a broad range of cell types to induce the expression of cytokines, chemokines, and metalloproteinases. As a result, secretion of IL-17A promotes inflammatory responses, which leads to the recruitment of neutrophils, enhancement of antibody production, and activation of T cells. Increased expression of IL-17A is seen in autoimmune diseases such as multiple sclerosis and rheumatoid arthritis. It is also associated with asthma, psoriasis, cancer, and transplant rejection.

Alternative namesInterleukin 17A, IL-17A, IL17A, Ctla-8, Ctla8, IL-17, Il17
Cell typeTh17
Gene ID16171
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