Application Note: Fluorescence technology is widely used to detect proteins in both the visible and near-infrared ranges. This product allows for superior signal detection and lower background noise when fluorochrome conjugated antibodies are used to visualize proteins in Western blotting and other applications. Antibody conjugates prepared with IRDye® 800 and IRDye® 700DX, Cy2™, Cy3™, Cy3.5™, Cy5™ and Cy5.5™, DyLight™405, DyLight™ 549, DyLight™ 649, DyLight™ 680, and DyLight™ 800 and Alexa Fluor® 488, Alexa Fluor® 532, Alexa Fluor® 546, Alexa Fluor® 647 and Alexa Fluor® 680 have been validated on various platforms using this product with superior results compared to other commercially available products. In the infrared range, where little to no autofluorescence occurs, specific signal is sharply evident from any background giving the best possible signal-to-noise ratio. This allows for detection levels in the picogram range which rivals the sensitivity of chemiluminescence on film for Western blotting. Superior results are also seen when this product is used for simultaneous labeling (multiplex) in Western blots or microscopy using various fluorochrome combinations for multicolor imaging. Membranes blocked with this product can be dried and are very stable. Membranes that are stored protected from light can be re-washed and/or rescanned.
Concentration Value: 1X
Western Blot Dilution: User Defined
General Disclaimer Note: 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 5199, Limerick, Pennsylvania, USA.
Physical State: Liquid (sterile filtered)
Purity and Specificity: Blocking buffer was prepared using ultra pure reagents dissolved in pharmaceutical grade water (WFI) and consists of a proprietary protein formulation in TRIS buffered saline at pH 7.6 with thimerosal added as an antimicrobial agent.
Background: This blocking buffer is specifically designed for Western blotting using fluorochrome conjugated antibodies. Pure nitrocellulose membrane is recommended for maximum performance. Other membranes, such as PVDF or nitrocellulose embedded in a support can be used, but may generate elevated backgrounds. Protein should be transferred from gel to membrane using standard protocols. Blocking buffer can be used for membrane blocking and to dilute both primary and secondary antibodies. Blocking Buffer for Fluorescent Western Blotting is suitable for use with imaging systems produced by Bio-Rad Laboratories, GE Healthcare, Alpha Innotech, FujiFilm Life Science, Licor Biosciences, UVP and Syngene.
Low Endotoxin: No
Other: Optimum performance is achieved using this product undiluted. However, this blocking solution can often be diluted 1:1 in TBS without a significant loss of performance.