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Gluten ELISA Kit

Gluten ELISA Test Kit is a double antibody sandwich enzyme-link immunoassay for the quantitative analysis of Gluten in feed, Rye, barley, wheat, rye, oats, their protein isolate or protein concentrated samples. 
Gluten ELISA Test Kit incorporates a fast, low-cost sample prep protocol with high recovery rates.

Sensitivity:20ppb
Specification: 96tests/kit

Description

Principle
Gluten ELISA Test Kit is a double antibody sandwich enzyme-link immunoassay for the quantitative analysis of Gluten in feed, Rye, barley, wheat, rye, oats, their protein isolate or protein concentrated samples. 
Gluten ELISA Test Kit incorporates a fast, low-cost sample prep protocol with high recovery rates.

An antibody with a affinity for Gluten is coated onto microwells. Standard or sample is added to the appropiate well and if Gluten is present it will bind to the coated antibody. Subsequently, Gluten bound to horse-radish peroxidase (HRP) is added and binds to the antibody not already occupied by Gluten present in the sample or standard. After this incubation period, the contents of the wells are decanted, washed and an HRP substrate is added which develops a blue color in the present of an enzyme. The resulting color intensity, has an inverse relationship with the target concentration in the sample. The levels of Gluten in the sample are determined by comparison of the color intensity of unknown samples with the standard curve, which is made using the Gluten standards provided in the kit.

Technical specifications 
Detect Limit : 20ppb
Specification:  96tests /kit
Reaction temperature: 37°C 
Detect time: 30min

Sample Preparation

- Rye, barley, wheat, rye, oats, their protein isolate or concentrated sample
1. Grind samples to 60 mesh sieve and thoroughly mix prior to sub-sampling.
2. Take 0.3g in 50mL centrifuge tube, add 30mL 1 × sample extractant.
3. Shake vigorously for 16 hours at 25°C.
4. Place for 2 min, centrifuge at 4000 rpm for 5 min.
5. Take the supernatant and dilute 70 fold with 1 × sample diluent, mix thoroughly.
   -100μL supernatant + 600μL Sample diluent, shake properly,
   - then take 100μL mixture, add 900μL Sample diluent, shake properly.

- Feed  
(which contains fermented wheat meal, extruded gluten less than 20%)
1.  Grind samples to 60 mesh sieve and thoroughly mix prior to sub-sampling.
2.  Take 2 g in 50mL centrifuge tube, add  0.2g NaCl, add 30 mL 1× sample extractant.
3.  Shake at 350 rpm vigorously for 2 hours at 25°C.
4.  Place for 10 min, centrifuge at 4000 rpm for 5 min.
5.  Take the supernatant and dilute 40 fold with the 1× sample diluent and mix thoroughly.
    - 50μL supernatant + 450μL Sample diluent, shake properly,
    - then take 100μL mixture, add 300μL Sample diluent, shake properly.

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