Depends on the saddle. Years ago I wore merino wool shorts with chamois, which wasn't really padding. But my saddle had a little padding, just enough to be comfortable for century rides and two-day rides of 200-250 miles.
One of my road bikes has a fairly stiff saddle with little padding, and I wear thicker padded shorts with that bike. I can ride that bike up to 20 miles with unpadded shorts, but it's not comfortable beyond that.
The other road bike has a fairly thickly padded saddle, as road saddles go, and I prefer Aero Tech Pro shorts, with a barely-there microfiber pad that's the closest thing I've found to old school chamois. It's been comfortable on long rides. But with thicker pads it's uncomfortable. Too much padding puts more pressure on the perineum and makes it worse.
Most comfortable pad I've tried? The 3D pads on inexpensive Przewalski shorts. Their shorts and bibs cost $15-$25 on Amazon and worth more than that just for the pads. The shorts themselves are pretty okay, about like the low priced Aero Tech Pro shorts. But the pads are exceptional. Thick, dense, very resilient and resistant to compression or losing loft, with an unusually smooth surface with dimples like golf balls. Comfortable on all day, all night rides. Hard to describe but it's terrific stuff. Wicks moisture well, yet never feels soggy. My only real gripe is my skinny thighs could use snugger fitting leg hems. But I have that same problem with all shorts and bibs. I need to buy longer leg shorts/bibs.