Universal Blocking Buffer (2X) for Western Blotting

Application Note: This is a 2X concentrated stock solution. Prepare a 1X working solution by diluting 1 part 2X concentrate with 1 parts TBS or equivalent. This product allows for superior signal detection and lower background noise in colorimetric, chemiluminescent, and fluorescent western blotting and other applications making it a convenient general purpose blocking agent. Antibody conjugates prepared with IRDye® 800 and IRDye® 700DX (Licor), Cy2™, Cy3™, Cy3.5™, Cy5™ and Cy5.5™ (GE Healthcare), DyLight™405, DyLight™ 549, DyLight™ 649, DyLight™ 680, and DyLight™ 800 (Thermo Fisher/Pierce) and Alexa Fluor® 488, Alexa Fluor® 532, Alexa Fluor® 546, Alexa Fluor® 647 and Alexa Fluor® 680 (Invitrogen/Molecular Probes) and ATTO (Atto-Tec) 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 Blocking Buffer can be dried and are very stable.

Western Blot Dilution: User Defined

Synonyms: multiplex Blocking Buffer, Immunoblot Blocking Buffer, Blocking Solution, Blocking Buffer Western Blot, Western Blot Blocking Buffer, Alexa Dye Blocking Buffer, DyLight Blocking Buffer, colorimetric, chemiluminescent blocking buffer, WB block

Background: It is an universal blocking buffer solution for Western Blotting. It is specifically designed for colorimetric, chemiluminescent, and fluorescent western blotting. Universal Blocking Buffer is recommended for blocking when phospho specific antibodies are used. 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. Western Blot blocking buffer is suitable for use with chemiluminiscent and fluorescent western blot imaging systems produced by Bio-Rad Laboratories, GE Healthcare, Alpha Innotech, FujiFilm Life Science, Licor Biosciences, UVP and Syngene.