Choosing the right biological buffer for the experiment can be tricky. This process is like playing "Guess Who". Like a game, you start with a very extensive list of buffers, and then slowly ask questions and research answers to narrow your choices. So far, you have learned a lot about Good's Buffers and are looking for more guides about this little game. In this article, we will introduce what you should consider when choosing a biological buffer. To
Step 1: Determine the pH of the experiment
In many experiments, you need to change the conditions to keep the protein in a satisfactory state in solution. For example, suppose you have performed an experiment that simulates biological conditions at pH 7.4, but your protein is unstable at this pH, so you need to optimize this situation. One thing to keep in mind, the target pH is usually about one pH unit (sometimes a little far away) from the isoelectric point (pI) of the protein. pI is the pH value at which a given molecule or protein has no net charge. When a protein reaches its pI, its solubility will not be so high.
Step 2: Ask the right questions
Just like a game, you must ask the right questions to ensure that you get the right buffer. Here are some important issues to focus on:
1. Is there any metal in the solution that may be complexed with the buffer?
2. Is your target protein an enzyme? Which category does it belong to?
3. What are the ionic strength requirements for ligand binding, protein stability or electrophoresis?
4. What literature tells me about the interaction of the proposed buffer with the system used?
All these questions will further help you find the most suitable buffer for the experiment. To
Step 3: Consider downstream experiments
One stumbling block that researchers often encounter is the discovery that the selected buffer is not suitable for downstream applications. For example, Tris is known to negatively affect Bradford analysis. Other biological buffers can negatively affect mass spectrometry. Perhaps you may have narrowed the scope of the buffer, only to find that it may not work well in the future. It's like a nightmare version of "Guess Who". In this case, you must do some research to find a suitable alternative. However, now that you have narrowed down the required features, it is easier to conduct this research.
Step 4: Find the right buffer
Great, now you know exactly what buffer to use. When it comes to purchasing, for most researchers, the reserve price and purity are basically important criteria for buying buffers. Of course, you can go to a more well-known company and consult the reputation of this company in the industry. Recommend a company here, Desheng not only provides excellent buffer products (TRIS, HEPES, BICINE, CAPS and other popular products). It also provides excellent content and service support to help you experiment. What's important is that the price is more competitive than other manufacturers.