Glutathione S-transferases (GSTs) are major detoxification enzymes which belong to Phase II defense enzymes; they can able to metabolize a variety of toxic chemical agents such as carcinogens, genotoxins, neurotoxins, and pesticides. Usually, GST will express when the living beings are encountered toxic chemical compounds. Acrylamide (ACR) is synthesized industrially and widely used in various industries. Usually, ACR formation occurs when food products prepared at high temperature. So that ACR is an environmental and food contaminant and it is well-proven neurotoxin. Due to highly mobile nature, birds that include poultry birds are main victims to xenobiotics (e.g., ACR) through food, water, and agricultural chemical formulas. In this study, ACR administered chick embryo brain GST activity level was assayed using 1-chloro-2,4-dinitrobenzene, and expression was assessed by western blot studies. The results show that the GST expression levels were increased in response to ACR by 24 and 48 h intervals. However, in 48 h interval, GST expression levels decreased slightly. Western blot studies also show similar pattern of GST expression. Immune blot studies showed similar GST band pattern as purification studies showed (our published work). In this study, enzyme activity and western blot analysis proved that the chick embryo brain GST was expressed more to detoxify ACR.