Place it at the exact point where a scene turns strange, then follow it with the Bruh Meme Sound for a slower second reaction. For a bigger build-up, open with It Was At This Moment He Knew before the reveal, then let this clip close the moment instead of adding another punchline.
About this sound
This is a tiny spoken clip labelled Psf5 Kava, lasting just over a second, with a clipped, hurried delivery that sounds odd out of context. It feels abrupt and slightly confusing, like a reply that arrives before anyone has finished thinking, which makes it funny precisely because it does not explain itself. That strange, sudden tone is why it works as a quick reaction.