Tissue/ cell preparation of Human T lymphocytes
Clone F10-89-4 reacts with all forms of CD45 expressed by all haematopoietic cells, except erythrocytes, having a higher level of expression on lymphocytes than on granulocytes.
Our Abpromise guarantee covers the use of ab30470 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 - 10 µg/ml.|
|Flow Cyt||1/10 - 1/50. ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.|
|IHC-Fr||1/500 - 1/1000.|
|IP||Use at an assay dependent concentration.|
|IHC-P||Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
Ab30470 staining CD45 in Jurkat cells. The cells were fixed with 4% formaldehyde (10min), 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 ab30470 at 5ug/ml (shown in green) and ab206369, Rabbit monoclonal to beta Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in red). This was followed by an incubation at room temperature for 1h with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed, at 1ug/ml (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 Jurkat cells fixed with 80% methanol (5 min).