We employed proximity labelling to derive a proteome of the luminal content of the Trypanosoma brucei endoplasmic reticulum (ER) and Golgi apparatus. We exploited the abundant ER chaperone BiP (Binding-immunoglobulin protein) as ER BioID bait and compare to a truncated version of BiP (termed BiPN) devoid of ER retention and thus trafficked to the Golgi apparatus, as well as parental controls. Our approach covers the two major life cycle stages of T. brucei, i.e., the procyclic form (PCF) and the bloodstream form (BSF). Additionally, we probed the inner nuclear membrane which can be viewed as a specialised ER domain separated by the nuclear pore, via the membrane integral nucleoporin NUP65 as BioID bait in PCF. Altogether, our combined approaches devise an inventory of the T. brucei secretory pathway that complements existing subcellular protein maps derived by proteomics and imaging.