Methods, systems and computer readable media for utilizing abstracted user-defined data to conduct network protocol testing are disclosed. According to one aspect, the subject matter described herein comprises a method that includes receiving, by a test system abstraction (TSA) module from a device under test (DUT), a packet containing a command that is associated with a native protocol and converting the command included in the received packet into a TSA protocol command. The method further includes processing the TSA protocol command at a TSA engine module that is provisioned with at least one instruction sequence enabling the TSA engine module to emulate a network test device and generating, by the TSA engine module, a TSA protocol command response in accordance with protocol behavior specified by the at least one instruction sequence.