Washing buffer is pipetted in and out of SPR ? unbound or nonspecifically bound sample is removed VIDAS Steps and Principle (cont. ) 3. Enzyme conjugated antibody (2nd) specific for target organism is pipetted in and out of SR ? 2nd antibody binds to organism captured previously by the 1st antibody VIDAS Steps and Principle (cont. ) 4. Washing buffer is pipetted in and out of SPR ? unbound or non-specifically bound 2nd antibody is removed VIDAS Steps and Principle (cont. ) 5. Enzyme substrate is pipetted in and out of the SPR substrate is broken down by enzyme on the 2nd antibody to form a fluorescent product Antibody enzyme conjugate Substrate VIDAS Steps and Principle (cont. ) 6. Reaction product is pipetted into the cuvette and the fluorescence read VIDAS system See lab sheet • Must be calibrated prior to use of each lot of kit
A standard is provided with the kit for the calibration – Calibrated value of standard is kept in the system for 14 days VIDAS result? (report shows 3 sets of number) 1. Background 2. RFV 3. TV 3 sets of number in VIDAS report 1. Background fluorescence in cuvette before the run 1. RFV (relative fluorescence value) = fluorescence in cuvette after the run minus the background 1. TV (test value) = RFV of sample / RFV of standard Analysis of VIDAS result • TV (test value) of sample is compared to that of threshold values – High and low threshold values – Pre-established by the supplier of the VIDAS system for the assay Analysis of VIDAS result TV of sample ? high threshold value < high threshold value ? low threshold value < low threshold value Result Interpretation + ? ? –
The Term Paper on The Effects of Ph and Salinity on Enzyme Function
Metabolism is the totality of all of an organism’s chemical reactions. Chemical reactions occur due to enzymes, a substance which acts as a catalyst in driving chemical reactions in order to produce a desired product (Campbell and Reece, 2002). A catalyst is usually a protein; however, some catalytic molecules counter this generalization. A discovery made in the early nineteen- nineties revealed ...
