Anti-Annexin V/ANXA5 抗体 (ab14196)

ab14196 staining Annexin V/ANXA5  in mouse embryonic 15 day brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with paraformaldehyde, ermeablized with 0.5% Triton X-100 in PBS, blocked with 10% serum for 1 hour at 25°C and antigen retrieval was by heat mediation in citrate buffer, pH 6. The sample was incubated with primary antibody (1/200) for 16 hours at 4°C. An Alexa Fluor^® 594-conjugated donkey anti-rabbit polyclonal (1/700) was used as the secondary antibody.

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ab14196 staining cells from a mouse organ of Corti (whole mount) by ICC/IF. The tissue was PFA fixed and permeabilized in 0.5% Triton X-100 prior to blocking in 5% goat serum, 2% BSA for 1 hour at 25°C. The primary antibody was diluted 1/200 and incubated with the sample for 2 hours at 25°C.  A Cy3® conjugated sheep anti-rabbit IgG antibody, diluted 1/200 was used as the secondary. Alexa Fluor 488® - Phalloidin was used to label the actin cytoskeleton and DAPI was used to label nuclei.

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ab14196 staining cells from a human pancreatic cell line by ICC/IF. Cells were PFA fixed and permeabilized in 0.1% Triton X-100 prior to blocking in 1% BSA for 30 minutes at 22°C.  The primary antibody was diluted 1/100 and incubated with the sample for 1 hour at 22°C. An Alexa Fluor® 488 conjugated donkey anti-rabbit antibody, diluted 1/1000, was used as the secondary.

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IHC image of Annexin V/ANXA5 staining in Human skin melanoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab14196, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.