Where should you put a tremolo pedal in your effects chain?
In the modulation section, after gain and before ambience.
Though some would argue that tremolo and vibrato pedals are their own effects category, we'd place them in the modulation category. This would dictate that they be positioned after gain effects (distortion, overdrive, etc.) but before ambient effects (delay and reverb).

A lot of guitar amps actually have tremolo built into their circuit, which makes its positioning a lot simpler. You don't have to worry about it at all.
But what about tremolo pedals?
Even if your amp has a built-in tremolo effect, a dedicated pedal is a lot more flexible and can give you some additional control over your sound. The same goes for a vibrato pedal which is similar to the tremolo sound. In fact, a lot of retailers actually group vibrato and tremolo into their own effects category.
Technically, your tremolo pedal should be placed after gain pedals, like distortion, and before ambient pedals, like delay or reverb.
And this is one of the problems that makes placing these effects so confusing. Because, technically, a tremolo pedal is a modulation effect.
A simple one - sure - but a modulation effect nonetheless.
The guy from JHS explains this really well.

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In a Straight Signal Chain (no effects loop)
Technically your tremolo pedal should be placed after gain pedals, like distortion, and before ambient pedals, like delay or reverb.
Here's a diagram of how it might look.

Tremolo pedals should be placed after distortion and before ambient effects, which are usually reverb or delay.
You should also consider some other more common modulation pedals, primarily the following:
- Chorus
- Phaser
- Flanger
These pedals are all in the modulation category and should be placed in the same position. How you place modulation pedals among each other doesn't really matter as much.
With an Effects Loop (send/return)
Your tremolo pedal is a good candidate for an effects loop, if your amplifier has one. This is usually labeled as a send/return on the back of your amp with an input and output jack.
Here's how it looks on the Mesa/Boogie Mark V:25 amp head:

Effects loops are an input and output jack on the back panel of your amp, usually labeled send and return. Putting your tremolo pedal in this loop is a good best-practice to follow.
Here's what that setup would look like:

If your amp has an effects loop, putting your tremolo pedal in that loop - outside of your main signal chain - is preferable.
How much does it really matter?
It's important to mention that effects pedal order and setup is always a matter of personal preference. Best practices can inform that process, but a lot of guitar players deviate from the norm depending on their rig and the style of music they play.
Sure - it matters to an extent that you at least know what the conventional wisdom is. But it's usually a breakable rule.
As with most things related to guitar rigs and setting up your gear, take our advice with a grain of salt.
Creative license that's informed by best practices is a good combination.
Conclusion
Since tremolo pedals are tricky to categorize, they're also difficult to place. Hopefully this has helped you sort through some of the confusion. Note that the best practices outlined here go for vibrato pedals as well. If you have questions, jump into the comments section below and we'll help out as much as possible.
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