From the time you hit start on the apparatus, the entire transfer takes about 7 minutes. The whole stack is then inserted horizontally into the rapid transfer apparatus. To perform the transfer, you apply your gel to a pre-assembled stack, which is then topped off with a flat electro-conductive layer. In this case, though, the layers come pre-assembled and soaked in buffer. The transfer stack is composed of the same layers used in a standard transfer (i.e. iBlot)Īnother common transfer method, often used by well-funded labs who can afford specialized equipment, is a semi-dry, or rapid, transfer. Because there is no conductive buffer in the areas that these bubbles are touching, no proteins will transfer at that spot, leading to a spotty or obscured signal on your membrane. However, if you’ve ever assembled one of these by hand, you know that it can be quite tricky to keep tiny air bubbles out from between the layers. Wet transfers are more quantitative and consistent than rapid blotting systems. This type of transfer can be run “hot” in an hour or two, but is often run slowly (from several hours to overnight) to preserve the crisp resolution of the bands. The transfer cassette is then immersed in a transfer apparatus filled with buffer, and an electrical current is applied to mobilize the proteins to migrate from the gel onto the membrane. These components are all well-soaked with transfer buffer, and then locked into a transfer cassette as a sandwich. To do a wet transfer, you put together a “sandwich” of layers, including filter paper, your gel, and the membrane. The basic wet transfer is probably the most common transfer method. ![]() In this article, we’ll talk about a few different ways to transfer a gel onto a membrane, and the advantages and disadvantages of both. Of course, it’s also finicky and slippery and prone to tiny pitfalls that can noticeably affect the quality of your final blot. I think that transferring Western blots is one the most enjoyable tasks to do in a lab: it’s quick, it’s messy, and on some gleeful level, it feels like a child’s art project gone wrong.
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