CD66 Monoclonal Antibody (CD66a-B1.1), Super Bright™ 702, eBioscience™

Product Specific Information

Description: The CD66a-B1.1 monoclonal antibody reacts with human CD66a, c, d, and e, which are also known as CEACAM1, CEACAM6, CEACAM3 and CEA, respectively. CD66 is expressed on endothelial and epithelial cells, as well as on neutrophils and monocytes. Myeloma and cancer cells also express CD66. Moreover, expression of this receptor can be induced on T cells, B cells, and CD16-negative NK cells. CD66 exists as multiple splice variants that differ in the length of their cytoplasmic domain, which contains an immunoreceptor tyrosine-based inhibitory motif (ITIM) that mediates the inhibitory functions of this cell surface protein on T and NK cells. CD66 is involved in cell adhesion and growth, neutrophil activation, and signaling. Finally, studies have shown that the CD66 variants interact with each other homo- and heterotypically.

Applications Reported: This CD66a-B1.1 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This CD66a-B1.1 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can 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 702 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 702 nm. We recommend using a 710/50 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: 702 nm; Laser: Violet Laser

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

Target Information

The CD66 molecules are 180-200 kDa glycoproteins of carcinoembryonic antigen family. They are present on all blood granulocytes and some tissue macrophages, but are absent from other hematopoietic cells. The expression of CD66 increases significantly on granulocytes upon their activation. CD66a (BGP1, CEACAM1) and CD66d (CGM1, CEACAM3), as well as CEACAM2 and 4, are transmembrane proteins, whereas CD66c (CEAL, NCA, CEACAM6) and CD66e (CEA, CEACAM5), as well as CEACAM7 and 8, are anchored to the plasma membrane by C-terminal glycosylphosphatidylinositol (GPI) lipid moiety.