Ly-6A/E (Sca-1) Monoclonal Antibody (D7), PE-Cyanine5, eBioscience™, Invitrogen™

Description

Description: The D7 monoclonal antibody reacts with mouse Sca-1, an 18 kDa member of the Ly-6 family of GPI-linked surface proteins. D7 reacts with both Ly-6E.1 and Ly-6A.2 molecules expressed by mouse hematopoietic stem cells, myeloid population and peripheral T and B cells. Non-activated splenocytes from Ly-6.2 strains including C57BL, SJL, 129, AKR, and others demonstrate higher frequency of D7-positive cells compared to the Ly-6.1 strains such as BALB/c, C3H, NZB and others. Sca-1 is expressed at high levels upon activation regardless of the Ly-6 haplotype. The Ly-6 family is involved in regulation and function of T cell activation. Sca-1 is a major phenotypic marker for mouse hematopoietic progenitor/stem cell subset. Applications Reported: This D7 antibody has been reported for use in flow cytometric analysis. Applications Tested: This D7 antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 0.125 μg per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Light sensitivity: This tandem dye is sensitive photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 μL cell sample + 100 μL IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 488-561 nm; Emission: 667 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser. Filtration: 0.2 μm post-manufacturing filtered. Ly-6A/E molecules are implicated in the regulation of T cell activation.