Product Description
Trans-Zeatin Riboside- ELISA Quantitation Kit
Product description
background | this ELISA assay utilise the principle of competitive binding to measure the concentration of hormone in plant extracts. The trans-zeatin riboside specific 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
Assay parameters
unspecific binding | 2.5 %
midrange(B/Bo=50%) | 0.02-10 pmol; detection limit | 12.25 pg (7 fmol)
linear range of measurment | 15-500 fmol; intraassay variability | 3.6 %
interassay variability | 4.3 %; amount of tracer per assay | 10 ng
