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.

Brilliant Violet™ 650 (BV650) is a tandem dye that emits at 649 nm and is intended for use on cytometers equipped with a violet (405 nm) laser. Please make sure that your instrument is capable of detecting this fluorochrome.

When using two or more Super Bright, Brilliant Violet™, Brilliant Ultra Violet™, or other polymer dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401-42) or Brilliant Stain Buffer™ (Product # 00-4409-75) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer or Brilliant Stain Buffer for more information.

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 # 00-5333-54) 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.

Our internal testing suggests that Brilliant Violet™ 650 (BV650) is not compatible with methanol-based fixation.

Excitation: 407 nm; Emission: 649 nm; Laser: Violet Laser.

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