Western blots are complex and offer many opportunities for something to go wrong. Can you tell what went wrong with this blot?
What happened with this blot is a little obscure and is something you may or may not have seen before. Pictured is a blot that had bacterial growth in the antibodies. Bacterial contamination could come from many sources in your protocol, including antibodies, blocking buffer, or equipment, although with good lab practices it’s unlikely.
How can I avoid bacterial contamination?
Bacterial contamination can be avoided by following good lab practices. Ensure that you do not reuse secondary antibodies or blocking buffer. Thoroughly clean all equipment, including incubation boxes and wash boxes, according to manufacturer recommendations.
Why does bacterial contamination matter?
As you might expect, bacterial contamination indicates a serious problem with a Western blot. Because the presence of bacteria calls the entire blot into question, a contaminated blot cannot be used for analysis or quantification. If your blot has been contaminated by bacteria, you will need to find the source of contamination and redo the blot.
For more troubleshooting examples and Western blotting tips, check out Good Westerns Gone Bad: Tips to Make Your Fluorescent Western Great.
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