Anti-Human Milk Fat Globule 1 antibody [SPM291] (ab17787)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Human Milk Fat Globule 1 antibody [SPM291] (ab17787)

ab17787 Milk Fat Globule Membrane Protein staining in human tissue. The primary antibody was diluted 1/50, and incubated for 30 minutes at room temperature.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Human Milk Fat Globule 1 antibody [SPM291] (ab17787)

IHC image of ab17787 staining in human normal breast formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM 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 ab17787, 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.

  Flow Cytometry-Anti-Human Milk Fat Globule 1 antibody [SPM291](ab17787)

Overlay histogram showing MCF-7 cells stained with ab17787 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab17787, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. 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 MCF7 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.