Anti-CD90 / Thy1 antibody [MRC OX-7] - Hematopoietic Stem Cell Marker (ab225)

Overlay histogram showing PC12 cells stained with ab225 (red line).

The cells were fixed with 4% paraformaldehyde (10 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 (ab225, 0.1µg/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, mouse IgG1 [B11/6], ab91353, 1µg/1x10⁶ cells) used under the same conditions.

Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. 

Ab225 gave a positive result in 80% methanol (5 min) fixed PC12 cells used under the same conditions.

ab225 staining CD90 / Thy1 in PC12 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 ab225 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150121, Goat polyclonal Secondary Antibody to Mouse IgM - mu chain (Alexa Fluor^® 488) 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.

Immunocytochemistry/ Immunofluorescence analysis of horse adipose and bone marrow stem cells labeling CD90 / Thy1 with ab225 at 1/1000 dilution. Cells were fixed in paraformaldehyde and permeabilized with Triton x100 0.1%. The cells were blocked with 10% BSA for 30 minutes at 37°C, followed by staining with ab225 at 1/1000 for 12 hours in PBS+IGEPAL+BSA+10%NGS at 4°C. Goat F(ab')2 Anti-Mouse IgG - (Fab')2 (Biotin), pre-adsorbed (ab5886) was used as the secondary antibody at 1/400 dilution.

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Immunocytochemistry/ Immunofluorescence analysis of rat sciatic nerve schwann cells and fibroblasts labeling CD90 / Thy1 with ab225 at 1/400 dilution. Cells were fixed in paraformaldehyde and permeabilized with 0.5% Triton X-100. The cells were blocked with 5% serum for 1 hour at 21°C, followed by staining with ab225 at 1/400 in 0.2% BSA for 1 hour at 37°C. A polyclonal goat anti-mouse Alexa Fluor^® 546 secondary antibody was used at 1/500 dilution.

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Rat CD90/Thy1 is N-glycosylated at three sites, giving rise to molecules with a range of molecular masses (25-37 kDa).