Histamine ELISA kit

Histamine ELISA Kit (ab213975) is a competitive ELISA kit for the quantitative determination of histamine in human, mouse, rat and canine sample matrices. This competitive, ultra-sensitive ELISA does not require acylation of histamine and has a time-to-answer of 2 hours. Please read the complete kit insert before performing this assay.

In this assay samples or standards are added to wells coated with a goat anti-rabbit IgG antibody. A polyclonal antibody to histamine and a solution of biotinylated histamine tracer are both added to the wells and the plate is incubated. During this incubation, the antibody binds the histamine in the sample or tracer in a competitive manner. The plate is washed, leaving only bound histamine on the plate. A solution of Horseradish Peroxidase conjugated Streptavidin (SA-HRP) is added to all wells and the plate is incubated. The plate is washed to remove excess conjugate. TMB substrate is added to the wells and the plate is incubated. Stop solution is added to stop the substrate reaction. The resulting yellow color is read at 450nm. The amount of signal is inversely proportional to the level of Histamine in the sample.

Neat (undiluted) methanol-extracted human serum and plasma samples, neat urine and tissue culture media in addition to mouse, rat and canine samples have been validated for use in this. However, due to variation in samples, dilution may be required. For detailed methanol-extraction method, please refer to Protocol Booklet.

Histamine is an endogenous short-acting biogenic amine synthesized from the amino acid histidine and is widely distributed throughout the body. Histamine is an important mediator of immediate-type-allergic reactions. Normally, there is a minimal amount of histamine circulating in the human body. An allergic reaction can occur when a substance enters the body (i.e. food, chemicals, insect bites, oils from plants). In response to the molecule, the body will then release larger quantities of histamine, which starts a cascade of events and begins an immune response. The body balances the histamine release by producing Epinephrine (adrenaline) which can help modulate the effects of histamine. When histamine is released, part of the cascade of events is inflammation. Antihistamines work by blocking the action of histamine and the resulting inflammation, amongst other symptoms, to provide relief.

Anaphylaxis occurs when there is a hypersensitive response. Reactions can range from mild to severe and, in extreme cases, can be fatal. Symptoms of hypersensitivity to histamine include hives, tingling feeling in the mouth, difficulty breathing, stomach cramping and diarrhea.

Elevations in plasma and tissue histamine levels have been reported during anaphylaxis and experimental systems investigating the allergic responses of the skin and airways. Because of its potent role in the immune response, histamine in the body is present in a very transitory state and can only be measured within minutes of release. The half-life of histamine in a biological system is four minutes before conversion to n-methyl histamine. N-methyl histamine is a major product of histamine metabolism, is present in urine and has a longer half-life than that of histamine.

Cross Reactivity

Histamine is also in a class of neurotransmitters called "Small Molecule Neurotransmitter Substances". This group includes molecules such as Serotonin, Epinephrine and Dopamine. Additionally, histamine plays a role in gastric acid secretion, assisting in the induction of acid production.