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Chemicals
Calcium chloride dihydrate (CaCl2.2H2O)
Citric acid monohydrate (C2H8O2.H2O)
Disodium hydrogen phosphate anhydrous (Na2HPO4)
Magnesium chloride hexahydrate (MgCl2 6H2O)
'Merthiolate'
(sodium ethylmercurithiosalicylate-'Thimerosal' - Sigma T5125)
Phosphate-citrate buffer Tablets (Sigma P4809)
Potassium chloride (KCl)
Potassium dihydrogen phosphate anhydrous (KH2PO4)
Sodium azide (NaN3)
Sodium bicarbonate anhydrous (NaHCO3)
Sodium carbonate anhydrous (Na2CO3)
Sodium chloride (NaCl)
Sodium hydroxide (NaOH)
Sulphuric acid (conc) (H2SO4)
Carbonate/bicarbonate buffer capsules (Sigma C3041)
Gelatine
Glycerol
Orthophenylene diamine dihydrochloride (OPD) or
OPD Tablets 30 mg (Sigma P8412)
Phenol red powder
Sand (must be sterile, washed, acid-treated, fine mesh sifted, laboratory grade)
Skimmed milk powder (Marvel or equivalent)
Tween 20 (Polyoxyethylene sorbitan monolaurate)
Biological Reagents
Coating sera
Rabbit antisera against each FMDV serotype and SVDV.
Each rabbit antiserum can be diluted 1:5 with PBS containing
either 0.2M sodium azide solution or 0.02% merthiolate
solution. Working stock antiserum reagent is stored at +4ºC.
Detection sera
'Blocked' guinea pig sera against each FMDV serotype
and SVDV mixed with equal volume of bovine serum and
heat inactivated at 56°C for 30 min (WRL, Pirbright, UK).
Each 'blocked' guinea pig antiserum is further diluted 1:5 in
(i.e., 1:10 final dilution) PBS containing either 0.2M sodium
azide solution or 0.02% merthiolate solution. Working
stock antiserum reagent is stored at 4ºC.
Conjugate
Peroxidase conjugated rabbit immunoglobulins to
guinea pig immunoglobulins (P141, Dako Ltd). Conjugate
is blocked with an equal volume of non-immune bovine
serum, diluted 1:5 with PBS and mixed with an equal volume
of sterile glycerol (i.e., 1:10 final dilution), and stored at 4ºC.
Reference control antigens
Aziridine inactivated cell culture supernatant fluids
containing each of seven serotypes of FMDV and SVDV.
Each control antigen is mixed with an equal volume of glycerol
and stored at -20ºC.
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