Post-translational modifications are known to intricately regulate protein functions, and among them, lysine acetylation has emerged as a particularly dynamic regulatory mechanism. Lysine acetylation involves the addition of an acetyl group to lysine residues, impacting protein structure, stability, and interactions . In recent years, this PTM has garnered a ention for its profound influence on cellular processes, especially in the context of key transcription factor complexes such as nuclear factor-kappa B. NF-κB is central to immune and inflammatory responses, helping with the process of cellular homeostasis. Dysregulation of NF-κB has been identified in numerous diseases, including cancer, highlighting the need for a comprehensive understanding of the factors influencing its activity. The addition of acetyl groups to lysine residues within NF-κB proteins introduces structural and functional changes that can intricately modulate the complex's activity. This can influence its DNA-binding affinity, altering its subcellular localization, and regulating its interactions with other proteins involved in the transcriptional machinery . These modifications have profound implications for the transcriptional output of NF-κB, dictating the expression of genes pivotal for cellular responses to stress, infection, and disease, including cancer . In cancer, distinct subunits of NF-κB are active, with RELB predominating in hematological cancers and RELA (p65) and/or c-REL being active in solid cancers. These subunits contribute to cancer formation and invasiveness through unique transcriptional activities . Similarly, in chronic inflammatory diseases, NF-κB p65-mediated transactivation plays a crucial role, and targeting this pathway has shown promise in mitigating disease progression. One approach to modulating NF-κB activity is through acetylation. Acetylation of NF-κB p65 has been shown to influence its transcriptional activity and cellular localization . This targeted acetylation may lead to altered NF-κB transcriptional activity, affecting downstream gene expression involved in cancer progression and inflammation. The acetylation of NF-κB p65can influence various aspects of NF-κB activity, including modulation of transcriptional activation, alteration in DNA binding affinity, modification of assembly with I-κBα, and changes in subcellular localization . These multifaceted effects highlight the potential of modulators of NF-κB signaling, offering new avenues for therapeutic intervention in cancer and chronic inflammatory diseases. To probe the regulatory landscape of lysine acetylation within NF-κB, we introduce a synthesized tool, Aspirin-NHS Ester. Aspirin contains an acetyl group that can be readily transferred to nucleophilic sites on proteins, such as lysine residues, through acylation reactions . Standalone aspirin has been implicated in its ability to acetylated lysine residues . By introducing a reactive NHS (N-Hydroxysuccinimide) ester group to aspirin, Aspirin-NHS is designed to enhance the specificity and efficiency of lysine acetylation within the NF-κB complex. This chemical modification ensures controlled and selective acetylation of lysine residues, minimizing off-target effects and enabling precise manipulation of NF-κB acetylation. Our goal is to contribute a specific tool that enables controlled modifications, fostering a deeper understanding of lysine acetylation's influence on the NF-κB complex and, by extension, its relevance in cancer-related processes. In this study, the choice of β-Conglycinin, a soy protein, as the model system is integral to our investigation into the effects of lysine acetylation within the NF-κB complex. β-Conglycinin consists of three subunits-α, α', and β-which collectively provide a complex yet well-defined environment for studying lysine acetylation. The abundance of lysine residues within the protein structure makes it particularly amenable to investigations focusing on post-translational modifications . By employing β-Conglycinin, we aim to capture the intricate interactions and consequences of lysine acetylation on individual subunits and, consequently, on the overall structural and functional dynamics of the NF-κB complex. This chosen model system not only facilitates a detailed exploration of the lysine-rich environment but also provides a practical framework for extrapolating findings to the broader biological context, enhancing the translational relevance of our study in the field of cancer research and therapeutic interventions.