Histamine Dynamic kit
Histamine quantification from Blood, PBMCs and cells
Histamine Dynamic kit HTRF®
The Histamine HTRF kit is a highly specific assay for quantifying an allergic response in whole blood and cell-based conditions.
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Ready-to-use
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Highly accurate
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Faster and more convenient than ELISA
Overview
Histamine (1H-imidazole 4-ethaneamine) is a biogenic amine known to play a key 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. Bacterial decarboxylation of the amino acid histidine in fresh fish produces histamine, which can be used as a health hazard biomarker indicative of bacterial contamination. The histamine kit accurately measures histamine from PBMCs and cell-based formats.
Benefits
- HIGHLY ACCURATE
- FASTER ALTERNATIVE TO ELISA OR WB
- READY-TO-USE KIT
Assay principle
The kit is based on a competitive format involving a specific antibody labelled with Cryptate (donor) and histamine coupled to d2 (acceptor). Histamine produced by cells competes with d2-labelled histamine for binding to monoclonal anti-histamine Eu Cryptate.
Assay Protocol
The histamine assay features a streamlined protocol with only two incubation steps:
- Sample with acylation buffer and acylation reagent
- Histamine detection using HTRF reagents.
This protocol requires a single, three-hour incubation period following the acylation step.
Key features
| 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 detection upon allergen stimulation
Four independent blood samples (Donor A to D) were used to measure the spontaneous release of histamine, as well as 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
Histamine detection upon REBL-2H3 stimulation
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 30 minute cell stimulation at 37 ºC using the standard HTRF histamine assay protocol.
Histamine detection upon PBMC stimulation
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 receptor with anti-IgE. Histamine detection was performed using the standard HTRF histamine assay protocol following 30 minutes of stimulation of peripheral blood basophils at 37ºC. Error bars represent a standard deviation of 3 replicates.
HTRF assays for Oncology and Inflammation
Signaling in the Immune System - Brochures
Product Insert Histamine Kit / 62HTMDPET-62HTMDPEG
62HTMDPET-62HTMDPEG - Product Insert
All your HTRF assays in one document! - Catalog
A guide to Homogeneous Time Resolved Fluorescence
General principles of HTRF - Guides
How HTRF compares to Western Blot and ELISA
Get the brochure about technology comparison. - Brochures
Compound allergenicity evaluation by measuring histamine release on whole blood
Predict compound toxicity in early drug discovery - Application Notes
Safety Data Sheet (DEU) Histamine Kit / 62HTMDPEG
62HTMDPEG - Safety Data Sheet
Safety Data Sheet (ELL) Histamine Kit / 62HTMDPEG
62HTMDPEG - Safety Data Sheet
Safety Data Sheet (FRA-FR) Histamine Kit / 62HTMDPEG
62HTMDPEG - Safety Data Sheet
Safety Data Sheet (ITA) Histamine Kit / 62HTMDPEG
62HTMDPEG - Safety Data Sheet
Safety Data Sheet (SPA) Histamine Kit / 62HTMDPEG
62HTMDPEG - Safety Data Sheet
Safety Data Sheet (ENG-GB) Histamine Kit / 62HTMDPEG
62HTMDPEG - Safety Data Sheet
Safety Data Sheet (ENG-US) Histamine Kit / 62HTMDPEG
62HTMDPEG - Safety Data Sheet
Safety Data Sheet (DEU) Histamine Kit / 62HTMDPET
62HTMDPET - Safety Data Sheet
Safety Data Sheet (ELL) Histamine Kit / 62HTMDPET
62HTMDPET - Safety Data Sheet
Safety Data Sheet (FRA-FR) Histamine Kit / 62HTMDPET
62HTMDPET - Safety Data Sheet
Safety Data Sheet (ITA) Histamine Kit / 62HTMDPET
62HTMDPET - Safety Data Sheet
Safety Data Sheet (SPA) Histamine Kit / 62HTMDPET
62HTMDPET - Safety Data Sheet
Safety Data Sheet (ENG-GB) Histamine Kit / 62HTMDPET
62HTMDPET - Safety Data Sheet
Safety Data Sheet (ENG-US) Histamine Kit / 62HTMDPET
62HTMDPET - Safety Data Sheet
Plate Reader Requirement
Choosing the right microplate reader ensures you’ll get an optimal readout. Discover our high performance reader, or verify if your lab equipment is going to be compatible with this detection technology.
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