Alexa Fluor® 647 Anti-ENO1 antibody [EPR19758]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 647 Anti-ENO1 antibody [EPR19758] (AB303584)

Immunofluorescence staining of ENO1 in a section of formalin-fixed paraffin-embedded human clear cell carcinoma*.

Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH9.0) using retrieval settings of 110°C for 40 minutes. The section was then incubated at room temperature for 1 hour with ab303584 at 1/100 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium^®.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

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

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-ENO1 antibody [EPR19758] (AB303584)

ab303584 staining ENO-1 in HeLa (human cervix adenocarcinoma epithelial cell line) cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3 M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab303584 at 0.04 µg/ml (shown in red) and ab195883 (Alexa Fluor®488 Anti-Tubulin antibody [YOL1/34]) at 5 µg/ml (shown in green). Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-ENO1 antibody [EPR19758] (AB303584)

Flow cytometry overlay histogram showing HeLa cells stained with ab303584 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab303584) (1x 106 in 100μl at 0.04μg/ml (1/12500)) for 30 min at 22°C.

Isotype control antibody (black line) was Rabbit IgG (monoclonal) Alexa Fluor® 647 (ab199093) used at the same concentration and conditions as the primary antibody. Unlabeled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 40 mW Red laser (638 nm) and 660/10 bandpass filter.

This antibody gave a positive signal in Hela cells fixed with 4% formaldehyde (10 min) / permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.