The Annexin 5 gene spans 29 kb containing 13 exons, and encodes a single transcript of approximately 1.6 kb and a protein product with a molecular weight of about 35 kDa.The protein encoded by this gene belongs to the annexin family of calcium-dependent phospholipid binding proteins some of which have been implicated in membrane-related events along exocytotic and endocytotic pathways. Annexin 5 is a phospholipase A2 and protein kinase C inhibitory protein with calcium channel activity and a potential role in cellular signal transduction, inflammation, growth and differentiation. Annexin 5 has also been described as placental anticoagulant protein I, vascular anticoagulant-alpha, endonexin II, lipocortin V, placental protein 4 and anchorin CII. Polymorphisms in this gene have been implicated in various obstetric complications.

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Validation of Annexin V knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with Annexin V antibody and analyzed using CytoFLEX.

Flow cytometric analysis of Annexin V expression in HT-1080 cells using Annexin V antibody . Green, isotype control; red, Annexin V.

Immunocytochemical staining of HT-1080 cells with Annexin V antibody . Nuclei were stained blue with DAPI; Annexin V was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: High. Scale bar, 20 μm.

Western blotting analysis using Annexin V antibody . Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with Annexin V antibody and HRP-conjugated goat anti rabbit secondary antibody respectively. Image was developed using FeQ™ ECL Substrate Kit .

Western blotting analysis using Annexin V antibody . Annexin V expression in wild type (WT) and Annexin V shRNA knockdown (KD) HeLa cells with 30 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with Annexin V antibody and HRP-conjugated goat anti rabbit secondary antibody respectively. Image was developed using FeQ™ ECL Substrate Kit .

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