Anti-A2B5 antibody [105]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-A2B5 antibody [105] (AB53521)

ab53521 staining A2B5 in primary rat neurons/glia, DIV14 (prepared from E18 rat hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. SDHEP) cells. The cells were fixed with 4% paraformaldehyde (10 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 ab53521 at 1µg/ml and ab6046. Cells were then incubated with ab150121 at 1/1000 dilution (shown in green) and ab150080 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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-A2B5 antibody [105] (AB53521)

ab53521 staining A2B5 in primary mouse neurons/glia, DIV14 (prepared from E18 mouse hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. C57EHP) cells. The cells were fixed with 4% paraformaldehyde (10 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 ab53521 at 1µg/ml and ab6046. Cells were then incubated with ab150121 at 1/1000 dilution (shown in green) and ab150080 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.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-A2B5 antibody [105] (AB53521)

ICC/IF image of ab53521 stained PC12 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab53521, 1μg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor^® 488 goat anti-mouse IgM (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor^® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue).

• ICC/IF

AbReview13490****

Immunocytochemistry/ Immunofluorescence - Anti-A2B5 antibody [105] (AB53521)

A2B5 antibody [105] - Stem Cell Marker (ab53521) used in immunocytochemical detection on Cor1 (mouse) neuronal stem cells. Cor1 neuronal stem cells were fixed in formaldehyde, permeabilized, blocked in 1% BSA for 30 mins at RT. ab53521 was incubated at 1/500 for 16 hours in TBS/BSA/azide/0.5% Triton. Secondary Antibody : anti mouse IgM conjugated : to Alexa Fluor^® 488 (1/1000).

This image is courtesy of an Abreview submitted by Mr Carl Hobbs

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-A2B5 antibody [105] (AB53521)

Overlay histogram showing SH-SY5Y cells stained with ab53521 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab53521, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-mouse IgM (mu chain) (ab97007) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgM [ICIGM] (ab91545, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.