Key facts
Detection method:Colorimetric,
Sample types:Urine, Cell culture media, Serum, EDTA Plasma,
Reacts with:Mouse, Rat, Human,
Assay type:Competitive,
Sensitivity:= 0.068 ng/mL,
Range:0.098 - 25 ng/mL,
Assay time:2h 30m,
Assay Platform:Pre-coated microplate (12 x 8 well strips)
Product details:
Histamine ELISA kit ab213975 is a competitive ELISA kit to measure histamine ( 2-(4-imidazolyl)ethylamine, HIS) with a sensitivity of 0.068 ng/mL.
How the assay works
Histamine (HIS)ELISA kit ab213975 is a competitive ELISA. There are 2 types of competitive ELISAs:
antigen down
: The sample antigen competes with a reference antigen for binding to a specific amount of labeled antibody. The reference antigen is pre-coated on a multi-well plate. The sample is added to the plate wells with a labeled antibody. Depending on the amount of antigen in the sample, more or fewer free antibodies will be available to bind the reference antigen. This means that the more of the antigen there is in the sample, the less reference antigen will be detected and the weaker the signal will be.
antibody down
: the labeled antigen and the sample antigen (unlabeled) compete for binding to the primary antibody, which is immobilized on the plate. The lower the amount of antigen in the sample, the stronger the signal due to more labelled antigen in the well.
Assay specificity
Our ELISA kits are rigorously validated to ensure the highest level of consistency and reproducibility.
Histamine ELISA kit ab213975 protocol summary
- Add standards or samples to the wells, incubate 2 hrs
- Wash, then add 50 µl of Biotinylated Antibody per well. Incubate 1 hr
- Wash, then add 50 µl of SP Conjugate per well. Incubate 30 min
- Wash, then add 50 µl of Chromogen Substrate per well. Incubate 20 min
- Add 50 µl of Stop Solution per well. Read at 450 nm immediately
Neat (undiluted) methanol-extracted human serum and plasma samples, neat urine and tissue culture media in addition to mouse and rat 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
+------------------------+----------------------+ |
Compound
Cross Reactivity
| +------------------------+----------------------+ | Histamine | 100% | +------------------------+----------------------+ | Nτ- methyl Histamine* | ≤ 2% | +------------------------+----------------------+ | Nα-Methyl Histamine* | ≤0.4% | +------------------------+----------------------+ | N-Acetyl Histamine | ≤ 0.01% | +------------------------+----------------------+ | L-Histidine | ≤ 0.02% | +------------------------+----------------------+
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.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-+4°C, Appropriate long-term storage conditions-+4°C, Storage information-+4°C
