Anti-mouse TNF-α mAb (MT11B10), biotin

Anti‑mouse TNF‑α mAb (MT11B10), biotin

3511-6-250

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Content

Monoclonal antibody MT11B10, 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 TNF-α in immunoassays such as ELISpot, FluoroSpot, and ELISA.

Serum/Plasma samples

Recommendation

MT11B10 is recommended as detection mAb in ELISpot, FluoroSpot, and ELISA in combination with capture mAbs MT1C8/23C9 (product code 3511-3).

Product details

ProductAnti-mouse TNF-α mAb (MT11B10), biotin
ApplicationELISpot, FluoroSpot, ELISA
AnalyteTNF-α
AntibodyMT11B10
ConjugateBiotin
ClonalityMonoclonal
ImmunogenRecombinant mouse TNF-α
HostRat
IsotypeIgG2a
ReactivityMouse
SpecificityNative and recombinant mouse TNF-α
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 MT11B10, 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.

Highlighted research, Published October 11, 2024

Immunoassays for vaccines
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.
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TNF-α

Analyte description

Tumor necrosis factor (TNF), also known as TNF-α, is produced by many different cell types, e.g., monocytes, macrophages, T cells, and B cells. Among the many effects of TNF-α are protection against bacterial infection, cell growth modulation, immune system regulation, and involvement in septic shock.

Alternative namesTumor necrosis factor-α, TNF-α, TNF-alpha, TNF-a, TNFa, Tumor necrosis factor-alpha, TNF, DIF, TNFSF2, TNFalphaa, Tnfsf1a, Tnlg1f, Tnf
Cell typeT cell, Tc, Th1, Th2, Th17, Tfh, Monocyte/MΦ
Gene ID21926
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