Automating the western blocking process.

Western blotting is a 40-year-old semi-qualitative process for identifying and visualizing proteins. Although this procedure is one of the most reliable methods for protein-related research, barely enough has been done to refine the process and improve efficiency and accuracy until recently.

Some advancements in the field of western blotting include automation, gel-free systems, one-step probing, etc. in this article, we will focus on how the automated western blot process improves the procedure.

Western blocking in a nutshell.

• Immunoblotting involves eight basic steps:
• Sample preparation.
• Gel electrophoresis to separate protein based on size and isoelectric point.
• Protein or membrane transfer.
• Blocking with bovine serum albumin or non-fat dry milk to prevent non-specific binding on the membrane.
• Incubation with the primary antibody.
• Incubation with the secondary antibody that binds with the primary antibody.
• Detection.
• Analysis.

Problems with manual western blocking.Blotting is very technical and sensitive and requires a high level of skill. Given there are multiple processes involved, there is a higher chance of error during handling. Apart from errors that occur during blocking, washing, and incubatiom, , other factors can affect the quality of the results like the concentration, reagent quality, materials, etc. For example, not letting the membrane dry after the protein transfer phase (some scientists wrongfully do this to save time) can lead to signal loss.

Some of the problems arising from the issues mentioned above include low target protein intensity, indiscreet or multiple bands, high background, and non-specific binding.

The implication of all this is that inexperience, fatigue, and poor reagents and materials can significantly affect the accuracy or sensitivity of the final results. Unnecessary repetitions waste time and money.

There are too many variables involved in the procedure, and reducing them to the barest minimum is crucial for obtaining accurate results. Therefore, the need for innovative technology arises. One solution to the challenges encountered during western blotting is automation.

How the automated process provides solutions.

An automated western blot processor lets you do more than just save time, it eliminates variation in fluid delivery, incubation, and washing. The device remembers up to 100 programmable protocol steps and can save 20 of them in memory. This increases reliability and precision with each step and delivers reproducibility.

Some of the benefits of using an automated western blot processor include:

Simplifies the process: The processor simplifies the whole procedure. Now, researchers or scientists work more effectively and don’t have to spend as much time as their counterparts who use the traditional process.

Reduces the margin for error: the automated blotting process reduces the variables involved compared to the traditional procedure, meaning there are few instances where mistakes can occur due to operator handling.

Saves time: As mentioned earlier, blotting is an arduous activity, and many of the steps involved are highly sensitive to time, temperature, storage conditions, vibration, etc. It is needless to say that an automated western blot process saves time, thereby giving scientists the opportunity to carry out other tasks, reduce fatigue and increase productivity.

Sponsor Ads

• Ad Space Available - Rent it Now!

Report this Page

Created on Jul 27th 2022 05:42. Viewed 279 times.

Share on APSense

Comments