Sample Extraction: HAVen extraction solution contains ethanol, gelatin & tannin binding reagent. HAVen also produces this solution stabilised & ready-to-use in an Improved Gluten Extraction kit, which saves both time and money; the inclusion of yellow dye in pre-filled extraction tubes helps confirm that samples have been diluted prior to testing.
In our laboratories, the use of either of these solutions is significantly superior to the conventional 40% ethanol:water extraction solution in terms of recovering gluten from raw materials and processed foods; the use of 40% ethanol:water is NOT recommended for the HAVen High Sensitivity Gluten ELISA (MORE INFO.)
Assay Reagents: The six Standards provided in each kit are colour coded and contain increasing amounts of red dye to aid the dilution step. Microwells are break-apart, allowing complete flexibility as to assay size. The enzyme reagent contains green dye as a pipetting aid. Two (breadcrumb) Controls are provided, a Low Level (>0<10mg/kg) and a Borderline Positive, at or above the proposed level for non-cereal derived foods (20mg/kg Gluten). The TMB Substrate used in the kit is particularly active and stable. Plates can be read for at least an hour after being stopped with low molarity acid. The kits contain no harmful preservatives e.g. azides/mercury compounds.
Assay Range: Protocol* 1. 5mg/kg Gluten to 100mg/kg Gluten (or higher) range.
Protocol* 2. 10mg/kg Gluten to 200mg/kg Gluten (or higher) range.
* Six ready to dilute Standards are provided in the kit. The lower five (250-5,000ng/mL), when diluted 1/10 as per unknown extracts, correspond to protocol 1 and the upper five (500-10,000ng/mL), when diluted 1/20 as per unknown extracts, correspond to protocol 2.
Limit Of Detection (LOD): <0.5mg/kg Gluten (on release LOD is usually <0.25PPM)
** Defined as the concentration of gluten interpolated from the assay standard curve at a response level equal to the Zero Standard OD450nm plus three Standard Deviations, calculated using at least 12 replicates of zero, three assays and two operators.
