background | this ELISA assay utilise the principle of competitive binding to measure the concentration of hormone in plant extracts. The N6-benzyladenosinespecific antibodies are precoated to the surface of the reaction wells. The plant extract sample, containing an unknown amount of hormone, is mixed in the reaction well with a known amount of a tracer to react with a limited number of antibodies in the reaction wells. During incubation the hormone in the sample competes with the tracer for the antibody binding sites. Unbound hormone, tracer and plant extract are washed out of the reaction wells. Following substrate addition which reacts with a tracer bound to the antibody and produces a yellow-coloured product. The absorbance of the sample is converted to concentration of hormone by means of a standard curve which is produced by simultaneously treating standards along with the samples.

reaction wells | antibody coated and blocked, 5pcs for 480 assays, 60 strips with 8 wells

tracer | 20 – 50 µl

tracer diluent | 5x250 mM TBS Tris, 10 mM NaCl, 1 mM MgCl2, pH 7.5 stock + 0.02 % NaN₃

reaction and wash solution | 10x TBS stock+0.02 % NaN₃

stopping reagent | 2x 5 N KOH stock

substrate diluent | 10x 500 mM NaHCO3 stock, pH 9.6+0 0.02 %. 0 0.02 % NaN₃

substrate | 100 mg

standards | 600 µl of each: 15.6 pmol, 7.8 pmol, 3.9 pmol, 1.95 pmol, 975 fmol, 488fmol, 244 fmol, 122 fmol, 61 fmol, 30.5 fmol, 15.2 fmol

assay development time | 4-5 hours

sensitivity | 0.01 to 10 pmol/50 µl

plant extract volume | 50 µl