Staining of C57BL/6 splenocytes with Anti-Mouse CD3e FITC (Product # 11-0031) and 0.03 µg of Rat IgG2a K Isotype Control PE-Cyanine7 (Product # 25-4321) (left) or 0.03 µg of Anti-Mouse Ly-6A/E (Sca-1) PE-Cyanine7 (right). Cells in the lymphocyte gate were used for analysis.
|Tested species reactivity||Mouse|
|Host / Isotype||Rat / IgG2a, kappa|
|Storage buffer||PBS, pH 7.2, with 0.1% gelatin|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C, store in dark, DO NOT FREEZE!|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||0.06 µg/test|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
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 0.06 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 to 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 of cell sample + 100 µL of 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: 775 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser
Ly-6A/E molecules are implicated in the regulation of T cell activation.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.