DyLight(TM) Multiplex 649/549 Duo Western Blot Kit

The DyLight conjugated secondary antibodies were prepared from monospecific antiserum by immunoaffinity chromatography using Rabbit or Mouse IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Goat Serum, Rabbit or Mouse IgG and Rabbit or Mouse Serum. No reaction was observed against Bovine, Chicken, Goat, Guinea Pig, Hamster, Horse, Human, Rat and Sheep Serum Proteins. These antibodies will react with heavy chains of rabbit or mouse IgG and with light chains of most rabbit or mouse immunoglobulins. Blocking buffer is specifically formulated to achieve superior reproducible western blotting images using this system. Wash buffer (10X TTBS) was aseptically filtered through a Millipore 0.22 micron filter into clean, pre-sterilized containers. The product was tested on trypticase soy agar for 24 hours, 48 hours and 72 hours and was found to be negative for bacteria. The The DyLight(TM) multiplex 649/549 Duo fluorescent Western blotting kit is suited for simultaneous detection and quantification of specific protein populations in a biological sample. Using a combination of two antibodies selected for minimal cross reactivity, fluorescent detection method enables simultaneous quantitative analysis of multiple proteins within the same sample on the same blot. The DyLight(TM) multiplex 649/549 Duo Western blot kit contain all the necessary components that are optimized for the simultaneous detection of multiple proteins on the same blot using DyLight(TM)-dye labeled secondary antibodies that are visualized in different fluorescence channels (649/549). The kit also includes blocking buffer, wash buffer, pre-stained protein standard and an incubation box for convinience and ultimate performance with minimal or no optimization. The fluorescent dyes such as DyLights(TM) when conjugated to secondary antibodies, offer a variety of benefits over traditional detection methods such as colorimetric and chemiluminescent detection. Multiplex detection using the correct lighting and filter conditions, enables the quantitation of multiple proteins and eliminates the need to strip and reprobe. Other benefits of fluorescent Western blotting include increased sensitivity, excellent signal stability over time as well as precise quantitative analysis with broader dynamic range and high linearity. Due to their exceptional photostability, DyLight(TM) dye conjugates can be archived and visualized several times without a decrease in signal.