Histamine Dynamic Kit

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. As part of an immune response to foreign pathogens, histamine is produced by basophils and by mast cells found in nearby connective tissues. Based on our patented HTRF® technology, the Histamine-Dynamic kit is a simple and sensitive approach for assessing immunogenicity reactions in whole blood and cell-based conditions.

Assay principle

The HTRF histamine-dynamic assay is an immunocompetitive assay which uses a d2-labeled histamine and an anti-histamine Cryptate-labeled antibody. The protocol involves an acylation of the Histamine from the standards and samples, to increase the assay sensitivity.

Product specifications

Standard curve 0.19 - 3,000 nM
Limit of detection (LoD) 0.21 nM
Assay range (LoD - IC90) 0.21 - 156 nM
Specificity: Acylated histamine 100%
Acylated L-histidine < 0.00067%
Acylated serotonin < 0.0067%

 

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

A plasma sample diluent (Ref # 62DLPDDD) can be purchased separately for plasma and serum applications.

Product performances

1. Allergenicity assessment in blood samples

Four independent blood samples (Donors A to D) were used to measure the spontaneous release of histamine, as well as releases upon stimulation with 5 allergen mixes including 6 grass mix, house dust mite, cat epithelium, inhalant mix, and common ragweed allergen.

*The anti IgE antibody (Ref # MABETS07) used as a positive control in the experiment can be purchased separately.

2. Histamine detection in rat basophils

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.

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

3. Histamine released by PBMCs

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.

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

Part#, inserts & MSDS

Ordering Info

DescriptionCat. noProduct insertMSDS
Histamine - Dynamic kit - 500 tests62HTMDPEG
Histamine - Dynamic kit - 96 tests62HTMDPET
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Companion products

DescriptionCat. noProduct insertMSDS
Diluent 2 - 20 ml62DL2DDD
Plasma sample diluent - 15 mL62DLPDDD
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Histamine - Dynamic kit - acylation reagent62HTMDADA
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Histamine - Dynamic kit - standard62HTMDCDA
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Histamine - Dynamic kit - acylation reagent diluent62HTMDDDA
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Histamine - Dynamic kit - acylation buffer62HTMDFDB
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Monoclonal antibody anti-IgEMABETS07
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