See kit insert for complete instructions.
Infrared-labeled antibodies and controls provided in this kit are sufficient to run approximately 25 immunoblots each measuring 10 cm x 10 cm. The amount of antibody supplied when diluted as recommended in our protocol will yield 1000 ml of working solution. Adjustments in volumes for larger or smaller blots will effect the number of blots detected. Blocking and washing buffers are provided in limited quantities so that the user can immediately begin to process immunoblots. These buffers can be re-ordered in larger quantities for economical use (see ''Replacement Parts'').
This kit allows for the individual (1-color) or simultaneous (2-color) detection of primary rabbit antibodies using an IRDye®800 conjugated secondary antibody (green) and primary mouse antibodies using an IRDye®700DX conjugated secondary antibody (red). The user is required to provide all primary antibodies. After protein separation and transfer, the membrane is blocked to prevent non-specific protein binding using a specially formulated blocking buffer for infrared fluorescent immunoblot detection. After washing the membrane, user-provided unlabeled primary antibodies are bound to the antigen. Additional washings are followed by the addition of IRDye®800 and IRDye®700DX conjugated secondary antibodies (see Figure 1). Final washing removes excess reagents and the membrane is further prepared for simultaneous scanning at two wavelengths to detect antibody complexes using a two channel infrared scanner such as LI-COR's Odyssey® Infrared Imaging System. The labeled antibodies are detected directly on the membrane, eliminating film, darkrooms, and messy substrates, while preserving high sensitivity (see Figure 2).
This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 326, Gilbertsville, Pennsylvania, USA.