{
  "url": "https://openlegislation.uk/eu-regulation-2010-206/annex-I/part-6/division-5/division-A/division-2",
  "json_url": "https://openlegislation.uk/eu-regulation-2010-206/annex-I/part-6/division-5/division-A/division-2.json",
  "legislation": "eur/2010/206",
  "legislation_title": "Commission Regulation (EU) No 206/2010 of 12 March 2010 laying down lists of third countries, territories or parts thereof authorised for the introduction into the European Union of certain animals and fresh meat and the veterinary certification requirements (Text with EEA relevance)",
  "legislation_url": "https://openlegislation.uk/eu-regulation-2010-206",
  "path": "annex/I/part/6/division/5/division/A/division/2",
  "kind": "division",
  "number": null,
  "heading": "Material and Reagents:",
  "text": "1.. Appropriate ELISA microtitre plates.\n2.. Antigen: supplied as a cell extracted concentrate, prepared as described below, and stored at either – 20 °C or – 70 °C.\n3.. Blocking buffer: phosphate buffered saline (PBS) containing 0,3 % BTV negative adult bovine serum, 0,1 % (v/v) Tween-20 (supplied as polyoxyethylene sorbiton monolaurate syrup) in PBS.\n4.. Monoclonal antibody: 3-17-A3 (supplied as hybridoma tissue-culture supernatant) directed against the group-specific polypeptide VP7, stored at - 20 °C or freeze-dried and diluted 1/100 with blocking buffer before use.\n5.. Conjugate: rabbit anti-mouse globulin (adsorbed and eluted) conjugated to horseradish peroxidase and kept in the dark at 4 °C.\n6.. Chromogen and substrate: Orthophenylene diamine (OPD-chromogen) at a final concentration of 0,4 mg/ml in sterile distilled water. Hydrogen peroxide (30 %w/v-substrate) 0,05 % v/v added immediately before use (5μl H 2 O 2 per 10 ml OPD). ( Handle OPD with care - wear rubber gloves - suspected mutagen ).\n7.. 1 Molar sulphuric acid: 26,6 ml of acid added to 473,4 ml of distilled water. ( Remember Acid must be added to water, never water to acid .)\n8.. Orbital shaker.\n9.. ELISA plate reader ( the test may be read visually ).",
  "extent": "E+W+S+N.I.",
  "status": null,
  "version": "current",
  "annotations": [
    {
      "type": "X",
      "label": "X1",
      "text": "Substituted by Corrigendum to Commission Regulation (EU) No 206/2010 of 12 March 2010 laying down lists of third countries, territories or parts thereof authorised for the introduction into the European Union of certain animals and fresh meat and the veterinary certification requirements (Official Journal of the European Union L 73 of 20 March 2010)."
    }
  ],
  "outstanding_effects": [],
  "provenance": {
    "source": "legislation.gov.uk",
    "source_url": "https://www.legislation.gov.uk/eur/2010/206/annex/I/part/6/division/5/division/A/division/2",
    "retrieved_at": "2026-11-01T14:50:53.580499+00:00"
  },
  "licence": "Open Government Licence v3.0",
  "licence_url": "https://www.nationalarchives.gov.uk/doc/open-government-licence/version/3/",
  "attribution": "Contains public sector information licensed under the Open Government Licence v3.0. Source: legislation.gov.uk, The National Archives."
}