ROTI®Quant universal, for protein quantitation,, for biochemistry, Density (D) 1,1 g/cm³, Storage temp. +15 to +25 °C, UN-Nr. 3082 Colorimetric test for detecting and quantifying soluble proteins. Universally applicable. Equals the well-known BCA assay Very easy to handle Highly sensitive measurement up to 0.5 µg protein Linear measuring range from 0.5 to 200 µg protein Can be used in 1-ml-scales and in microtiter plates (can be automated) Measurement with standard filters (492 nm wave length) or at 503 nm wave length Accelerated reaction and increased sensitivity in mild heat The special reagent formula effects a high tolerance against interfering factors such as detergents, alcohol and buffer salts, thus making the kit a universal tool for measuring proteins in almost every solution. Mechanism The basic mechanism, a PCA enhanced Biuret reaction, is identical with the one used for the well known BCA protein assay. Its result, however, is a bright blue colour which may be measured with standard filters of approx. 500 nm wave length.ROTI®Quant universal is based on a biuret reaction in combination with a highly specific colorimetric enhancer reaction. The Cu++ ions of reagent 2 are reduced to Cu+ ions in the alkaline reaction media of the test. The colour development of the copper-specific enhancer reaction is directly proportional to the concentration of the protein which is present.The resulting purple-coloured copper complex exhibits a strong light absorption with a maximum of 503 nm, although the wide maximum of the absorption curve also permits highly sensitive and no-loss measurements at 492 nm. The reaction does not form a true end point measurement: the colour develops slowly at room temperature, this, however, can be accelerated in heat. High sensitivity, therefore, can be achieved even after a short time. Mechanism The kit contains 500/200 ml ROTI®Quant universal reagent 1 (Art. No. 0118) and 40/16 ml ROTI®Quant universal reagent 2 (Art. No. 0119).Contents of this Kit may not be bought separately. Fig. A: ROTI®Quant universal, standard curve with BSA (bovine serum albumin). Blank-value corrected. Measurement of 100 µl each protein solution. Fig. B: Independence of ROTI®Quant-universal-assay of detergents and other additives, examples. Blank-value corrected. Measurement of 100 µl each protein solution.(With kind permission of Dr. Angelika Böttger, Zoological Institute, Ludwig-Maximilians-University Munich) Fig. D: Independence of ROTI®Quant-universal assay of SDS. Microassay in microtiterplates (60 min, 37 °C). Technical Information Detection Biuret reaction/PCA (equals BCA) PDF Technical Documentation (PDF)
