Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling CD44 with ab238464 at 0.085µg/ml, followed by a ready to use secondary. Membranous staining on rat spleen is observed. The section was incubated with ab238464 for 5 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use secondary.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C6 (rat glial tumor glial cell) cells labeling CD44 with ab238464 at 0.854µg/ml, followed by ab150113 AlexaFluor^®488 Goat anti-Mouse secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining on C6 cell line. The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab179504 Anti-beta IV Tubulin antibody - Microtubule Marker at a 1/200 dilution followed by ab150080 AlexaFluor^®594 Goat anti-Rabbit secondary at a 1/1000 dilution.
The negative controls are as follows:
-ve control 1: ab238464 at 1/500 dilution, followed by ab150080 AlexaFluor^®594 Goat anti-Rabbit secondary at 1/1000 dilution.
-ve control 2: ab179504 at 1/200 dilution, followed by ab150113 AlexaFluor^®488 Goat anti-Mouse secondary at 1/1000 dilution.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized rat splenocytes labeling CD44 with ab238464 at 0.854µg/ml, followed by ab150113 AlexaFluor^®488 Goat anti-Mouse secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining on rat splenocytes. The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab179504 Anti-beta IV Tubulin antibody - Microtubule Marker at a 1/200 dilution followed by ab150080 AlexaFluor^®594 Goat anti-Rabbit secondary at a 1/1000 dilution.
The negative controls are as follows:
-ve control 1: ab238464 at 1/200 dilution, followed by ab150080 AlexaFluor^®594 Goat anti-Rabbit secondary at 1/1000 dilution.
-ve control 2: ab179504 at 1/200 dilution, followed by ab150113 AlexaFluor^®488 Goat anti-Mouse secondary at 1/1000 dilution.

Flow cytometric analysis of C6 (Rat glial tumor glial cell) cell line labeling CD44 with ab238464 at 0.474µg/ml (red) compared with a Mouse monoclonal IgG Isotype Control (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti mouse IgG (Alexa Fluor^® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.

Gated on viable cells.