Arsenic (As) contamination of the environment is a widespread problem with the situation at its worst in the South Asian region of West Bengal, India, and Bangladesh. In order to cope up with the problem and to be able to engineer plants in near future, there is a need to thoroughly understand the kinetics of As uptake and its impact on responses of plants. In this work, Brassica juncea (L.) Czern was used as a model system to understand kinetic interaction between As uptake and transport to the shoot and corresponding biochemical responses. The seedlings of B. juncea were exposed to 100 μM arsenite [As(III)] in hydroponics for different time points of 1, 4, and 24 hours. The As concentration was found to show a gradual increase in different tissues with time and the level of As followed the order: lower root greator than upper root greator than shoot greator than top leaves at all the times points. However, the level of hydrogen peroxide (H2O2) and the activities of NADPH oxidase, superoxide dismutase (SOD) and ascorbate oxidase (AO) were increased even at 1 h in lower shoot and leaves. The expression of transporters genes, NIP1;1, NIP2;1, NIP5;1 and NIP6;1 was found significantly up-regulated at 24 hours. This work establishes a kinetic relationship between As accumulation and ensuing biochemical responses in B. juncea.