Ab14581 staining human normal skin. Staining is localised to the nucleus.
Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

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

ab14581 staining Mitotic proteins - Mitosis Marker in Human Saos-2 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.25% Triton in PBS and blocked with 1% BSA for 1 hour at room temperature. Samples were incubated with primary antibody (1/500) for 1 hour. An Alexa Fluor® 594-conjugated Goat anti-mouse IgG polyclonal (1/250) was used as the secondary antibody.

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

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Flow Cytometry analysis using anti-phospho-Ser/Thr-Pro, MPM-2 (ab14581). HeLa cells were treated with colcemid (green), or untreated (grey).

ab14581 (green) labeling phospho-proteins in metaphase cells. The spindle, centrosomes and centromeric regions are stained. DNA is counterstained with propidium iodide (red).