How do I optimize my western blot experiments?

There are several ways one is able to optimize western blot experiments. 

• One method is utilizing a gel (for electrophoresis) with a high concentration for optimal separation of histone proteins, ensuring clarity in the resulting bands. This means selecting a gel with a higher polymer concentration, such as polyacrylamide. 
• Another way to optimize western blots is to experiment with multiple blocking buffers to find the most suitable one for each antibody-antigen combination. It's important to note that milk-based blocking buffers are composed of substances like glycoproteins and biotin which may disrupt the signal. 
• The choice of membrane can influence the sensitivity of detection and the ability to bind proteins. Nitrocellulose and polyvinylidene difluoride (PVDF) are two common types of membranes used in Western blotting. 
• Additionally, selecting a buffer that suits the gel, the target protein, and membrane type is essential for successful western blotting. 
• One should always confirm the effectiveness of protein transfer by staining the membrane with a total protein stain or utilizing a pre-stained protein ladder. 
• Transfer conditions such as time, voltage and buffer composition should also be optimized according to the characteristics and size of the target protein. Larger proteins necessitate lengthier transfer times and lower voltages, whereas smaller proteins can be transferred faster using higher voltages. 
• Lastly, after selecting the appropriate antibody, the next step is to find the optimal working concentration. This involves testing various antibody dilutions while keeping other factors constant to achieve the best Western blot results.