Tissue/ cell preparation: cultured bone marrow cells.
ab53453 recognizes the Ly-6B.2 antigen. Ly-6B.2 is a ~25-30 kDa GPI-anchored, heavily glycosylated protein expressed on neutrophils, inflammatory monocytes and some activated macrophages. High levels of expression are seen in bone marrow, spleen, lung and lymph nodes. N-glycanase treatment of thioglycollate elicited peritoneal neutrophil lysates lowers the apparent molecular weight of Ly-6B.2 to ~15 kDa.
ab53453 has been successfully used for the immunomagnetic depletion of neutrophils during the enrichment of primitive hematopoietic cells from bone marrow and the depletion of myeloid cells in vivo.
Our Abpromise guarantee covers the use of ab53453 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-FoFr||Use at an assay dependent concentration. PubMed: 20038794|
|IHC-Fr||Use at an assay dependent concentration.|
|Flow Cyt||Use at an assay dependent concentration.
Use 10µl of the working dilution to label 106 cells in 100µl.
Immunohistochemical analysis of acetone-fixed frozen mouse spleen, labelling Neutrophil with ab53453 at a dilution of 1/100 incubated for 1 hour at 25°C in 5% goat serum in TBS. The secondary used (to amplify the signal) was a Goat anti-rat polyclonal Alexa Fluor® 488 conjugate at a 1/400 dilution.