Anti-human IL-1α mAb (MT513), biotin

Anti‑human IL‑1α mAb (MT513), biotin

3414-6-250

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Content

Monoclonal antibody MT513, 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 human IL-1α in immunoassays such as ELISpot, FluoroSpot, ELISA, and Western blot.

Serum/Plasma samples

Recommendation

MT513 is recommended as detection mAb in ELISpot, FluoroSpot, and ELISA in combination with capture mAbs MT641/642 (product code 3414-3). The antibody is also suitable for Western blot.

Product details

ProductAnti-human IL-1α mAb (MT513), biotin
ApplicationELISpot, FluoroSpot, ELISA, Western blot
AnalyteIL-1α
AntibodyMT513
ConjugateBiotin
ClonalityMonoclonal
ImmunogenRecombinant human IL-1α
HostMouse
IsotypeIgG1
ReactivityHuman
Specificity

Native and recombinant human IL-1α. The antibody binds to the pro-form and the active form of IL-1α.

Purification

Purified from in vitro cultures by protein G affinity chromatography.

Biotinylation

Biotinylated through reaction with a N-hydroxysuccinimide ester of biotin.

Concentration0.5 mg/ml
Supplied in

PBS with 0.02% sodium azide. Sterile-filtered (0.2 µm).

Contents

Monoclonal antibody MT513, biotinylated. Supplied at 0.5 mg/ml in PBS with 0.02% sodium azide.

Shipping and Storage

Shipping

Shipped at ambient temperature.

Storage

Store 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-1α

Analyte description

Interleukin 1α (IL-1α) is a pro-inflammatory cytokine primarily produced by monocytes, macrophages, and neutrophils. IL-1α induces acute phase responses such as fever, T cell, and macrophage activation. 

Alternative namesInterleukin 1α, IL-1α, hematopoietin 1, Interleukin-1alpha, IL-1 alpha, IL1α, Interleukin-1 alpha, IL-1A, IL1, IL1-ALPHA, IL1F1
Cell typeMonocyte/MΦ, mDC
Gene ID3552
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