Small volumes of anti-ANXA4 antibody vials may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid that has accumulated.

The ANXA4 antibody is involved in various processes such as cell proliferation, migration and invasion. Previous studies showed that ANXA4 expression was significantly elevated in human GBC tumors. High ANXA4 expression was associated with lymph node metastasis, invasion depth and TNM stage in patients with GBC (P 0.001).

To investigate the role of ANXA4 in GBC progression, we performed experiments using ANXA4-knockdown cells. ANXA4 mRNA and protein were down-regulated in GBC-SD and NOZ cells after shRNA mediated knockdown of ANXA4. Knockdown of ANXA4 resulted in reduced cell proliferation, migration and invasion. Additionally, cleaved caspase 3 and -9 were detected in the ANXA4-knockdown groups but not in the scrambled control groups, indicating that the ANXA4-knockdown effect was cell specific.

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ANXA4 mutant cells also showed impaired cell migration, invasion and adhesion. ANXA4 wild-type and mutant plasmids were transfected into THESCs, and the resulting stably-transformed cells were screened for ANXA4 expression by G418 selection. ANXA4 mutant clones were isolated and confirmed by WB. ANXA4 mutant cells were also tested for apoptosis by caspase activity assay and RT-qPCR.

Moreover, SWISS-MODEL analysis demonstrated that the p.G8D mutation of ANXA4 changed the 3D structure of the protein and weakened its interaction with p50 and IkBa. Therefore, ANXA4 p.G8D mutation might act as an oncogenic driver in GBC by interacting with the NF-kB signaling pathway.