Ly-6G/Ly-6C Monoclonal Antibody (RB6-8C5), PerCP-eFluor™ 710, eBioscience™, Invitrogen™

Description

Description: The RB6-8C5 monoclonal antibody reacts with mouse Ly-6G, a 21-25 kDa protein also known as the myeloid differentiation antigen Gr-1. A GPI-linked protein, Gr-1 is expressed by the myeloid lineage in a developmentally regulated manner in the bone marrow. While monocytes only express Gr-1 transiently during their bone marrow development, the expression of Gr-1 on bone marrow granulocytes as well as on peripheral neutrophils is a good marker for these populations. eBioscience testing indicates that in the bone marrow and lysed whole blood, the antibody clone RB6-8C5 also stains cells that express the highest levels of Ly6c (as defined by staining with antibody clone HK1.4). It is recommended that 1A8-Ly6G (cat. 9668) be used when looking at Ly-6G specific targets. Applications Reported: This RB6-8C5 antibody has been reported for use in flow cytometric analysis. Applications Tested: This RB6-8C5 antibody has been tested by flow cytometric analysis of mouse bone marrow cells. This may be used at less than or equal to 0.25 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. PerCP-eFluor 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome. 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 (Product # 00-8222-49) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-57) 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 nm; Emission: 710 nm; Laser: Blue Laser

Granulocytes are a category of white blood cells characterized by the presence of granules in their cytoplasm. They are also called polymorphonuclear leukocytes (PMN or PML) because of the varying shapes of the nucleus, which is usually lobed into three segments. In common parlance, the term polymorphonuclear leukocyte often refers specifically to neutrophil granulocytes, the most abundant of the granulocytes. Granulocytes or PMN are released from the bone marrow by the regulatory complement proteins.