Description: The CBRM1/5 monoclonal antibody reacts with an activation-specific epitope of human Mac-1. CBRM1/5 binds a subset of Mac-1 molecules on neutrophils and monocytes after stimulation with chemoattractants or phorbol esters but does not recognize Mac-1 on resting myeloid cells. Through interactions with its ligands, Mac-1 participates in adhesive cell interactions. The epitope recognized by this mAb localizes to the I domain on the alpha chain of Mac-1 very close to the ligand binding site in a region that is widely exposed. CBRM1/5 blocks Mac-1 dependent adhesion to fibrinogen and ICAM-1 and inhibits chemoattractant-stimulated adhesion of eosinophils to the Intercellular Adhesion Molecule-1 (ICAM-1). It should be noted that low level activation may occur during processing of freshly drawn blood. Therefore the CBRM1.5 antibody may exhibit some binding to Mac-1 in these unstimulated samples. However, higher levels of Mac-1 expression are observed in activated samples when compared to unstimulated cells.

Applications Reported: This CBRM1/5 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This CBRM1/5 antibody has been pre-diluted and tested by flow cytometric analysis of stimulated normal human peripheral blood cells. This may be used at 5 µL (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.

Super Bright 600 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 600 nm. We recommend using a 610/20 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome.

When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining 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) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 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: 405 nm; Emission: 600 nm; Laser: Violet Laser

Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.