Anti-MUC1 antibody [SM3]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MUC1 antibody [SM3] (AB22711)

IHC image of MUC1 staining in a formalin fixed, paraffin embedded human breast carcinoma tissue section*, performed on a Leica Bond™ 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 ab22711, 10 μg/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.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• IHC-P

AbReview20649****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MUC1 antibody [SM3] (AB22711)

ab22711 staining MUC1 in human breast carcinoma tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using 10mM citrate buffer. The primary antibody was used undiluted for 18 hours at 4°C. An undiluted HRP-conjugated goat anti-mouse IgG polyclonal was used as secondary antibody.

Image courtesy of an anonymous Abreview.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-MUC1 antibody [SM3] (AB22711)

ab22711 staining MUC1 in MCF7 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab22711 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-MUC1 antibody [SM3] (AB22711)

Overlay histogram showing MCF7 cells stained with ab22711 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab22711, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150117) at 1/2000 dilution for 30 min at 22°C.

Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab170190, 1μg/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-Triton X-100 for 15 min used under the same conditions.