During the pilot, which launches on 18 April, two escalators will be "standing only" during morning rush hour, with at least "at least one other" will be free for those who want to walk.
It follows initial tests late last year that TfL heralded as a "success", saying 30 per cent more people could travel on the escalator if they stood on both sides.
Peter McNaught, operations director at London Underground, said: “It may not seem right that you can go quicker by standing still, but our experiments at Holborn have proved that it can be true. This new pilot will help us find out if we can influence customers to stand on both sides in the long term, using just signage and information.
"Anyone who wants to walk on the other escalators will be free to do so, but we hope that with record numbers using the Tube, customers will enjoy being part of this experiment to find the most efficient ways of getting around.”
TfL noted that at stations "with very long escalators", such as Holborn, few customers choose to walk, meaning that much of left-hand-side of the escalator is unused.
A previous test held in November and December 2015, proved that as many as 30 per cent more customers could use an escalator in the busiest parts of the day if they stood on both sides.
But it seems the plans are largely going down like... a person standing on the left hand side of the escalator during rush hour.
@BBCr4today So banning people walking up escalator at Holborn will be quicker. Surely making EVERYONE walk would be even quicker.— Phil Walder (@WallyFlea) March 10, 2016
The @BBCr4today tells me that socialism has come to Holborn tube with compulsory standing, faster for all, slower for movers— Barbara Cookson (@filemot) March 10, 2016