Yes, you can add products from multiple sellers and checkout just one time, however, please be advised that if just one seller doesn't ship to your location, you won't be able to continue and place the order. When this is the case we recommend removing other seller products and trying again. Most US-based sellers will ship to all 50 states, so traditionally this won't be an issue.
Additionally, upon checkout, note that each seller sets a shipping rate, and those are combined for a total shipping charge for all sellers you are ordering from. If you feel the rate is high for what you plan to order, simply remove other seller's products and see what each seller's rates are one by one.