The high court refused to hear two appeals by Judge Roy Moore on Monday, who was suspended in August.
Moore suspension from work came after his refusal to comply with a federal judge's order to remove the granite monument because it violated constitutional church-state separation.
The judge, who was elected as Alabama's chief justice in 2000, has been accused of violating judicial ethics.
He is fighting his suspension, and a hearing has been set for 12 November in Montgomery, the state capital.
Court order removal
The 2.3 tonne stone marker was placed in the rotunda of the judicial building in Montgomery by Moore and his followers in 2001.
The monument was removed on 27 August after eight associate justices of the Alabama Supreme Court decided to comply with an order by US District Court Judge Myron Thompson.
It has been locked in a closet since then.
Moore, who has strong support from fundamentalist Christian groups for his championing of public displays of the 10 Commandments, vowed on Monday to continue his struggle in the court of public opinion.
"The United States Supreme Court may turn a deafening ear to the people of Alabama, but they will not be able to avoid the thundering roar that's going to be heard across this country," Moore said in a news conference outside Montgomery.
Moore's supporters also pledged to place the issue on the national agenda.
Civil liberties groups, who have accused Moore of imposing his personal religious views on others and using his office to promote the Christian faith, are equally adamant about resisting such attempts.
The controversial monument is
under lock and key
A number of civil liberties groups sued in 2001 on behalf of three Alabama lawyers who said they had been offended by the monument's display.
Thompson and then a US appeals court in Atlanta ruled the monument must be removed.
In one appeal, Moore's lawyers told the Supreme Court the federal judge lacked jurisdiction to issue an injunction to Alabama's chief justice requiring the monument's removal.
In the second appeal, his lawyers urged the high court to overrule its long-standing test dating back to 1971 on what represents an unconstitutional government endorsement of religion.
According to the original constitutional definitions, they said this monument would not represent a government "establishment of religion" forbidden by the First Amendment.
One group involved in the lawsuit, Americans United for Separation of Church and State, said the high court's action meant Moore has lost his final appeal and the legal controversy is over.
"This is the end of the legal line for Roy Moore," said the Rev. Barry Lynn, the group's executive director.
"It is time for Moore to face facts: he's on the wrong side of the Constitution. Religious symbols belong in our homes and houses of worship, not our courthouses."
In August, the Supreme Court rejected a request by Moore for a last-minute stay to prevent the monument's removal.
The high court in recent years has let stand other US appeals court rulings that have barred the display of large granite monuments engraved with the Ten Commandments on the grounds of state capitals.