Gentle animal–human interactions, such as stroking, can promote positive emotions and thus welfare in cattle. While previous studies showed that stroking at the ventral neck elicited the most positive reactions in cows, intra-specific allogrooming in cattle includes different body regions and is probably guided partly by the receiver. Thus, we compared heifers’ (n = 28) reactions to stroking with the experimenter either reactively responding to perceived momentary preferences of the heifers or exclusively stroking the ventral neck. Independently of the stroking style, longer durations of neck stretching and contact occurred during stroking, supporting our hypothesis of a positive perception of stroking. We did not confirm the predicted decrease in heart rate and increase in heart rate variability, but instead found a slightly increased mean heart rate during stroking. The different stroking styles elicited differences in the heifers’ ear positions: “reactive” stroking led to longer durations of low ear positions during stroking, while during “ventral neck” stroking, the duration of back up increased. However, no other behaviours differed significantly between different stroking styles, indicating that the exact manner of stroking applied in our treatments seemed to be less important in the promotion of positive affective states in cattle through gentle human–animal interactions.