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Histamine assay

Histamine is a small biogenic amine known to play an important role in immediate hypersensitivity and inflammation reactions as well as in several atopic diseases such as allergic rhinitis, bronchial asthma and eczema. Based on our patented HTRF® technology, the Histamine kit is a highly specific assay for quantifying an allergic response in cell-based conditions.

Features

  • Cost-effective alternative to ELISA
  • Homogeneous assay
  • Single plate protocol
  • Highly accurate
  • Amenable to automation

Applications

  • Histamine measurement
  • Allergy & Asthma assessment
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The HTRF histamine assay is an immunocompetitive assay which uses XL665-labeled histamine and an anti-histamine Cryptate-labeled antibody. The assay has a two-step procotol: acylation and detection. Acylation and detection can both be carried out in a single plate for simplified automation.

Detection limit: 1.8 nM histamine
Dynamic range 10 – 400 nM histamine
EC50 histamine 43 nM
Specificity: Acylated histamine 100%
Acylated L-histidine < 0.00067%
Acylated serotonin < 0.0067%

Histamine standard curve

Histamine standard curves were performed using the designated final assay volumes and plate types.

 

Rat basophils (RBL-2H3) were cultured in the presence or absence of 2.5 µM ionomycin to stimulate histamine release. Total histamine content was determined by lysing cells and spontaneous release was evaluated by incubating the cells in medium only.

Biomarker histamine performance

Histamine detection was performed following 30 minute cell stimulation at 37ºC using the standard HTRF histamine assay protocol.

 

Similar studies were performed using human peripheral blood basophils from two individual donors. Non-specific release of histamine using Ionomycin was compared to specific degranulation induced by cross-linking a membrane-bound IgE with anti-IgE. Error bars represent a standard deviation of 3 replicates.

Biomarker histamine performance 2

Histamine detection was performed using the standard HTRF histamine assay protocol following 30 minutes of stimulation of peripheral blood basophils at 37ºC.

Ordering Info

DescriptionCat. noProduct insertMSDS
Histamine kit - 1,000 tests62HTMPEB
Histamine kit - 20,000 tests62HTMPEC

Companion products

DescriptionCat. noProduct insertMSDS
Diluent 2 - 20 ml62DL2DDD
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Histamine acylation reagent - 1,000 tests62HTMADA
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Histamine acylation reagent - 20,000 tests62HTMADC
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Histamine calibrator 62HTMCDA
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Histamine acylation reconstitution buf. 1 mL62HTMFDA
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Histamine acylation reconstitution buf. 20 mL62HTMFDD
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Histamine control 62HTMTDA
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Reconstitution buffer 2 - 13 ml62RB2RDD
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