Consistent, Specific Protein Detection
Detect the presence of specific proteins in biological samples, measure changes in protein size, activation state, or posttranslational modifications, or investigate protein-protein interactions. Consistent Western blot success is possible. However, it is threatened by a variety of factors such as variability in sample preparation, inconsistent antibody performance, and the presence of background signal.
Buffers are essential at every step in the Western blot protocol to effectively denature proteins, maintain the proper pH, and block non-specific antibody binding. When selecting Western blotting buffers, you need reliable, customizable buffers to meet your needs. Boston BioProducts can help.
The Process
Following cell or tissue lysis, extracted proteins are denatured by sample buffer and then separated by size via polacrylamide gel electrophoresis (PAGE). Resolved proteins are transferred to a membrane and detected through binding of protein-specific primary antibodies followed by species-specific secondary antibodies conjugated to enzymes, fluorophores, or biotin for detection.
How we simplify the process
Western blotting success requires consistent and specific detection of target proteins with limited background signal. Though standard protocols and reagents will be suitable for many applications, protocol and reagent optimization is necessary for other immunoblot assays. Boston BioProducts offers a comprehensive range of reliable standard Western blotting buffers along with custom buffer services to ensure you achieve consistent results that propel your research forward.
Ways Boston BioProducts can help
Optimize your western blotting protocols with customized buffer services
Sometimes Western blot protocols are antibody-specific and require optimization. At Boston BioProducts, we overcome this challenge by providing a wide range of high-quality standard Western blotting buffers and flexible custom buffer solutions to ensure our customers get fit-for-purpose buffers that consistently work for all their immunoblot assay needs.
Assure reproducibility with lot-to-lot consistency
Consistency is key to reproducible research and accurate diagnostics. You've worked tirelessly to ensure reproducible results, combat user variability, streamline sample handling, and optimize protocols. The buffers you use should work just as hard to provide accurate, reliable results. Boston BioProducts western blotting buffers provide full traceability and lot-to-lot reproducibility, eliminating worry about changes in reagent performance.
Remain ready for experiments of all sizes with flexible sizing
A range of off the shelf Western Blotting buffer sizes and customized solutions from Boston BioProducts ensure that your needs are met, regardless of the scale of your experiment. Buffers are available in flexible sizing from a few mililiters up to hundreds of liters packaged in your choice of vials, bottles, carboys, or bioprocessing containers.
Remain western-ready with supply chain reliability
Limited buffer availability, lengthy turn-around times, and insufficient customer service and transparency can negatively impact your critical research by delaying your experiments. Boston BioProducts offers a reliable source of buffers and rapid delivery to your laboratory to keep your inventory stocked and your experiments moving forward.
Learn more about how
Boston BioProducts can help
Featured Products for Western Blotting
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Related Applications
FPLC Protein Purification
SDS PAGE
Frequently Asked Questions
The best blocking buffer will depend on your protein(s) of interest and the detection methods you are using. Bovine Serum Albumin (BSA) is more effective than nonfat milk for biotin-avidin systems. BSA is also suitable for detection of phosphorylated proteins but is not compatible with lectin probes.
Phosphate-Buffered Saline (PBS) is a widely used buffer for biological applications and frequently used for immunoassays. That is because PBS, composed of monobasic potassium phosphate, dibasic sodium phosphate, potassium chloride and sodium chloride, is an isotonic and non-toxic buffer that mimics the osmolarity and ion concentrations of the human body.
For the many applications, PBS and Tris-Buffered Saline (TBS) are interchangeable. However, when detecting phosphorylated proteins, TBS should be used instead of PBS so that the phoshpho-specific antibodies do not bind the phosphate group present in your PBS buffer.
A low concentration detergent in the washing step helps remove excess antibody. Tween-20, diluted in TBS or PBS in the range of 0.05% to 0.1%, is commonly used.