Buffers & Concentrates

SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis, describes a technique widely used in biochemistry to separate proteins according to their electrophoretic mobility (a function of the length of a polypeptide chain and its charge) and no other physical feature. SDS is an anionic detergent applied to protein sample to linearize proteins and to impart a negative charge to linearized proteins. 2X SDS-PAGE Sample Loading Buffer is suitable for laboratory involved in protein biochemistry. Visit our newly expanded web site at www.rockland.com for methods using this and other buffers.

RIPA (Radio-Immunoprecipitation Assay) Lysis Buffer enables rapid, efficient cell lysis and solubilization of proteins from both adherent and suspension cultured mammalian cells. It has long been a widely used lysis and wash buffer for small-scale affinity pull-down applications, such as immunoprecipitation, since most antibodies and protein antigens are not adversely affected by the components of this buffer. In addition, RIPA Lysis Buffer minimizes non-specific protein-binding interactions to keep background low, while allowing most specific interactions to occur, enabling studies of relevant protein-protein interactions.  The following RIPA Lysis Buffer components have the following effects: Tris-HCl is a buffering agent prevents protein denaturation, NaCl is a salt that prevents non-specific protein aggregation, NP-40 is a non-ionic detergent to extract proteins; Na-deoxycholate and SDS are ionic detergents to extract proteins; and sodium azide is a bacteriostatic agent added to re

Fluorescence technology is widely used to detect proteins. However, many common visible fluorophores often result in considerable background fluorescence in the visible range. Visible fluorophores are rarely used for membrane-based protein detection because of this high background. IRDye™800, IRDye™700DX, Alexa Fluor® 680 and Cy5.5™ antibody and reagent conjugates are specifically designed for protein detection methods that use longer-wavelength, near-infrared (IR) fluorophores to visualize proteins in western blotting and other applications. IRDye™800 and IRDye™700DX fluoresce outside the range of most autofluorescence and therefore specific signal is sharply evident from any background giving the best possible signal-to-noise ratio. Detection levels in the picogram range on Western blots rival the sensitivity of chemiluminescence on film. IRDye™800 conjugates are also suitable for immunofluorescence microscopy using commercially available excitation/emission filters in the 780nm/820n

BlockOut® Universal Blocking Buffer Solution for Western Blotting is specifically designed for colorimetric, chemiluminescent, and fluorescent western blotting. BlockOut® 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. BlockOut® can be used for membrane blocking and to dilute both primary and secondary antibodies. BlockOut® buffer is suitable for use with chemiluminescent and fluorescent western blot imaging systems produced by Bio-Rad Laboratories, GE Healthcare, Alpha Innotech, FujiFilm Life Science, Licor Biosciences, UVP and Syngene.