Anti-Annexin V/ANXA5 antibody [1F4-1A5] (ab54775)

ab54775 Western Blot analysis of ANXA5/ANXA5 expression in Hela cells.

This image was generated using the ascites version of the product.

Annexin V/ANXA5 antibody (ab54775) used in immunohistochemistry at 3ug/ml on formalin fixed and paraffin embedded human colon.

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ab54775 at 10 ug/ml staining Annexin V/ANXA5 in human Hela cells by Immunocytochemistry / Immunofluorescence.

This image was generated using the ascites version of the product.

Overlay histogram showing HeLa cells stained with ab54775 (red line). The cells were fixed with 100% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab54775, 1µg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

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Immunoprecipitation of ANXA5 transfected lysate using ab54775 monoclonal antibody and Protein A Magnetic Bead and immunoblotted with a ANXA5 rabbit polyclonal antibody.

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Western Blot analysis of ANXA5 expression in transfected 293T cell line by ab54775.
Lane 1: ANXA5 transfected lysate(35.9 KDa).
Lane 2: Non-transfected lysate.

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ab54775 staining Annexin V/ANXA5 in HeLa cells treated with pentoxifylline (ab120725), by ICC/IF. Increase of Annexin V/ANXA5 correlates with increased concentration of pentoxifylline, as described in literature.
The cells were incubated at 37°C for 24h in media containing different concentrations of ab120725 (pentoxifylline) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab54775 (10 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

This image was generated using the ascites version of the product.