Anti-mouse IL-2 mAb (5H4), biotin

Anti‑mouse IL‑2 mAb (5H4), biotin

3441-6-250

Publications: 110
Documents
Questions?
Validated for our assays.

Content

Monoclonal antibody 5H4, biotinylated. Supplied at 1 mg/ml in PBS with 0.02% sodium azide.

In stock

Delivery 4-9 business days

Shipping $0

Complementary products

Complementary products
1 / 1

Product specifications

Intended use

This monoclonal antibody enables specific detection of mouse IL-2 in immunoassays such as ELISpot, FluoroSpot, and ELISA.

Serum/Plasma samples

Recommendation

5H4 is recommended as detection mAb in ELISpot, FluoroSpot, and ELISA in combination with capture mAb 1A12 (product code 3441-3).

Product details

ProductAnti-mouse IL-2 mAb (5H4), biotin
ApplicationELISpot, FluoroSpot, ELISA
AnalyteIL-2
Antibody5H4
ConjugateBiotin
ClonalityMonoclonal
ImmunogenRecombinant mouse IL-2
HostRat
IsotypeIgG2b
ReactivityMouse
SpecificityNative and recombinant mouse IL-2
PurificationPurified from in vitro cultures by protein G affinity chromatography.
BiotinylationBiotinylated through reaction with a N-hydroxysuccinimide ester of biotin.
Concentration1 mg/ml
Supplied inPBS with 0.02% sodium azide. Sterile-filtered (0.2 µm).
ContentsMonoclonal antibody 5H4, biotinylated. Supplied at 1 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.

Highlighted research, Published October 11, 2024

Immunoassays for vaccine research
Explore the critical role of immunoassays like ELISpot and FluoroSpot in vaccine development, from evaluating immune responses to tracking vaccine efficacy and potency. These assays provide sensitive, single-cell insights for advancing research and development of viral, bacterial, and cancer vaccines.
Loading publications...

IL-2

Analyte description

Interleukin 2 (IL-2) is predominantly produced by activated T cells. IL-2 promotes the proliferation, differentiation, and survival of antigen-activated T cells and NK cells. After antigen stimulation, the cytokine also promotes the differentiation of T cells into effector and memory T cells. Therefore IL-2 plays an important role in the body's response to infection. IL-2 is also key to tolerance, as it promotes the differentiation of certain immature T cells into regulatory T cells in the thymus, ultimately preventing autoimmune diseases. 

Alternative namesInterleukin-2, Interleukin 2, IL-2, IL2, T cell growth factor, Il-2
Cell typeT cell, Th1, Th2, Th17, Tfh
Gene ID16183
1 / 2