In January 1941, the brigade was committed to the Western Desert Campaign, going into action for the first time around Bardia, which was captured from the Italians over several days. This was followed later in the month by the capture of Tobruk, where the 16th Brigade remained while the 6th Division's other two brigades, the 17th and 19th, took part in other actions around Derna and Benghazi. In March, the division concentrated around Amiriya as they prepared to deploy to Greece, in response to an expected German invasion. During the Battle of Greece, the 16th Brigade occupied Seria before establishing itself in the Veria Pass. As the German attack began, the 2/1st Battalion provided rearguard cover while the brigade withdrew alongside the New Zealand 4th Brigade. A German breakthrough at Tempe Gorge resulted in a further withdrawal during which the 2/2nd and 2/3rd Battalions provided rearguard security as Anzac Corps withdrew through Larissa. After this, the brigade was ordered to evacuate, and elements were withdrawn by sea at Kalamata. The majority of the brigade was evacuated back to Egypt and subsequently re-formed in Palestine; however, elements of the brigade landed on Crete. The 2/1st was landed as a whole and deployed to defend the airfield at Retimo, while elements of the 2/2nd and 2/3rd were formed into a 16th Brigade Composite Battalion. During the fighting, the 2/1st was largely destroyed during the brief Battle of Crete, with the majority being taken prisoner.