重组Alexa Fluor® 647荧光Anti-GM130抗体[EP892Y] - cis-Golgi Marker

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-GM130 antibody [EP892Y] - cis-Golgi Marker (AB195303)

Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling GM130 with ab195303. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% 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 ab195303 at 1/2500 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

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

This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5 min).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-GM130 antibody [EP892Y] - cis-Golgi Marker (AB195303)

ab195303 staining GM130 in HeLa cells. The cells were fixed with 100% methanol (5min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab195303 at 5μg/ml (shown in red) and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1/100 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with an AlexaFluor®488 Goat anti-Mouse secondary (ab150117) at 2μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

This product gave a positive signal in 4% formaldehyde (10min) fixed HeLa cells under the same testing conditions.

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

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-GM130 antibody [EP892Y] - cis-Golgi Marker (AB195303)

Overlay histogram showing HeLa cells stained with ab195303 (red line). The cells were fixed with 80% methanol (5 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 (ab195303, 0.1μg/1x10⁶ cells) for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (Alexa® 647) (0.2μl/1x10⁶ cells) for 30 min at 22°C. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a 25mW red solid state diode laser (635nm) and 675/30 bandpass filter.

This antibody gave a positive signal in HeLa fixed with 4% formaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 647 Anti-GM130 antibody [EP892Y] - cis-Golgi Marker (AB195303)

Immunohistochemical analysis of paraffin-embedded human cervix cancer tissue labeling GM130 with ab195303 at 1/100 dilution.

Membranous staining on human cervix cancer.

The primary antibody was incubated for 60 mins at room temperature (shown in red). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount®.

The immunostaining was performed on a Leica Biosystems BOND RX instrument.

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

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 40 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 647 Anti-GM130 antibody [EP892Y] - cis-Golgi Marker (AB195303)

Immunohistochemical analysis of paraffin-embedded human breast tissue labeling GM130 with ab195303 at 1/100 dilution.

Membranous staining on human breast.

The primary antibody was incubated for 60 mins at room temperature (shown in red). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount®.

The immunostaining was performed on a Leica Biosystems BOND RX instrument.

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

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 40 mins.