First time last week in Bank of America, I heard a teller tell the young fellow in front of me, "we don't have enough cash on hand to cash this check". He was a young blue collar kid, How much could it have been for, he sold his car. She wouldn't tell me. "I had to ask". Back to the question, if you have the cash in your account, you can take it out while it clears over night. Sometimes it's a time like 2 or 3 pm, but good rule of thumb is overnight. I just got one from a guy and it was pending, "that night" and cleared the following morning. That's the cool thing about online banking.