AOAC ERP Gluten Assays - Dec 2018
4. Safety β‐mercaptoethanol. – Cocktail (patented), necessary for sample preparation, contains β‐ mercaptoethanol. Use a chemical hood for sample preparation. Sulfuric acid . – Stop solution contains 1 N sulfuric acid. Avoid skin and eye contact. a) In the last week of August / first week of September, you will receive a shipment containing test kits, Cocktail (patented), practice samples, pre‐plates and test samples. b) The shipment will also contain a receipt acknowledgement form for you to return, indicating receipt and conditions of the items shipped. Please complete this form and email a scanned copy to Katharina Scherf ( k.scherf.leibniz‐lsb@tum.de ) on the same day the shipment is received. c) Test samples, practice samples, pre‐plates and test kits are to be stored at common refrigerated temperatures until assay. The Cocktail (patented) has to be stored at room temperature (20 °C – 25 °C). Prepare all solutions and reagents according to the method directions. d) All work should be completed by September 21 st , unless other prior rearrangements are made. If this schedule cannot be met, the Study Director should be notified as soon as possible. a) Sample extraction and assay procedure should be carried out in separate rooms if possible. It is recommended to do the extraction of the samples and the ELISA on the same day. b) Take the ELISA kit out of the fridge. All components should be at room temperature (20‐25 °C, 68‐77 °F) before use. Outside the test kit packaging this will take up to two hours, especially for the bigger buffer bottles due to their large volume. In case of any doubt, check temperature. c) Heat a water bath to 50 °C (122 °F). Check if the space in this water bath is big enough to contain 21 sample vials during extraction. The water level in the water bath should be at least as high as the 35 ml mark on the extraction vials to ensure sufficient temperature interchange. 5. Receipt and Storage of Materials 6. General preparations
d) In case of any doubt please contact the study director.
7. Prepare 80% ethanol according to the instructions for use (5.2)
a) Add 4 parts of pure ethanol (96%) to 1 part of distilled or deionized water e.g. add 560 ml ethanol p.a. to 140 ml distilled water and shake well. This will be enough 80% ethanol for the extraction of 21 samples.
8. Prepare the washing buffer according to the instructions for use (10.1)
a) The washing buffer is provided as a 10fold concentrate. Before use, the buffer has to be diluted 1:10 (1+9) with distilled water (i.e. 100 ml buffer concentrate + 900 ml dist. water).
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