AgriseraTMB Diluent (100 ml)

Instructions for a 96 well micro titer plate 1, Equilibrate the substrate to the assay temperature before use, 2, Complete all required incubations with antibodies and HRP labeled reagents, 3, Wash the plate at least 4 times with phosphate buffered saline or Tirs buffered saline containing 0,1% Tween-20, THen shake and blot all residual buffer from plate wells, 4, Add 100 µl of the TMB solution to each well and incubate 5-30 minutes, depending upon the activity of the HRP probe, If color develops too rapidly, zero order kinetics will not prevail, Dilution of antibody, or HRP-labeled reagent may be required, As alternative Agrisera TMB/Diluent pack can be used, 5, To stop the reaction use:-equial volume of 0,3 M sulfuric acid (100 µl), The yellow chromogen in the stopped reaction should be read at 450 nm, This will provide highest sensitivity of detection, -Monitoring the assay kinetics at may be done at 650 nm as a function of time, The reaction can be stopped to preserve the blue chromogen using an equal volume, 100 µl of 0,1% sodium fluoride or 0,15% sodium dodecyl sulfate, The blue chromogen in the stopped reaction should be measured at 650 nm, Adjustments of time, reagent volume and temperature require standardization by the end user, In case of any further questions please contact us

One component ready to use reagent. It contains 2.5 mMol L-1 TMB and Hydrogen Peroxide in a buffer at pH 3.6. It also contains non-toxic stabilizers and is recommended to be used tto adjust sensitivity of Agrisera TMB HRP Substrate.

Stable for 3 years at +2-8° C. It can also be stored at room temperature up to six months, and can also be frozen. Protect the substrate from exposure to sunlight.