At Woof we’re big on constantly measuring and testing. As such, we regularly test different versions of web pages and email newsletters, in order to improve conversions.
We split tested (also known as A/B testing) the subject line of a recent email newsletter that we sent out for a client to a selected sample of their subscriber list. Check out the email open rates below:

Thing is, if we’d have gone with gut instinct, we’d most definitely have chosen version A! It seemed to be the more punchy of the two. Just goes to show.
We then sent the remaining few thousand newsletters using the version B subject line, of course!
Always, always measure and test, everywhere you can.



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Just to add to what Dave said here.
I was shocked to discover that not only did the the subject line affect the number of people opening the email. It actually appeared to affect the level of interaction within the email. The number of click throughs from people that opened campaign B were almost double that of campaign A. The content was exactly the same.
Did people trust the content of the email more because they liked the subject line? Who knows. But it is a very powerful case for split testing.