To achieve successful assistance with long-horizon web-based tasks, AI agents must be able to sequentially follow real-world user instructions over a long period. Unlike existing web-based agent benchmarks, sequential instruction following in the real world poses significant challenges beyond performing a single, clearly defined task. For instance, real-world human instructions can be ambiguous, require different levels of AI assistance, and may evolve over time, reflecting changes in the user's mental state. To address this gap, we introduce RealWebAssist, a novel benchmark designed to evaluate sequential instruction-following in realistic scenarios involving long-horizon interactions with the web, visual GUI grounding, and understanding ambiguous real-world user instructions. RealWebAssist includes a dataset of sequential instructions collected from real-world human users. Each user instructs a web-based assistant to perform a series of tasks on multiple websites. A successful agent must reason about the true intent behind each instruction, keep track of the mental state of the user, understand user-specific routines, and ground the intended tasks to actions on the correct GUI elements. Our experimental results show that state-of-the-art models struggle to understand and ground user instructions, posing critical challenges in following real-world user instructions for long-horizon web assistance.