Staining of LPS-stimulated BALB/c splenocytes with Anti-Mouse CD11b Pacific Blue® and Rat IgG2a κ Isotype Control eFluor® 605NC (cat. 93-4321) (left) or Anti-Mouse CD86 (B7-2) eFluor® 605NC (right). Total viable cells were used for analysis.
Contents: Anti-Mouse CD86 (B7-2) eFluor® 605NC Catalog Number: 93-0862 Formulation: aqueous buffer, 0.09% sodium azide, may contain carrier protein/stabilizer Storage Conditions: Store at 2-8°C. Light sensitive material. This product is guaranteed for 6 months upon receipt when stored properly. Clone: GL1 Host/Isotype: Rat IgG2a, κ
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93-0862-41
25 tests
93-0862-42
100 tests
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*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µL to 5 µL. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.
Description
The GL1 monoclonal antibody reacts with mouse CD86, an ~80 kDa surface receptor also known as B7-2. CD86 & CD80 are members of the B7 family of costimulatory molecules. CD86 is expressed at low level on B cells, macrophages, and dendritic cells and is upregulated on B cells through a variety of surface stimuli including the BCR complex, CD40 and some cytokine receptors. CD86 is also expressed by activated mouse T cells and thioglycolate-elicited peritoneal cells. In addition to CD80 (B7-1), CD86 is a counter-receptor for the T cell surface molecules CD28 and CD152 (CTLA-4). This interaction plays a critical role in T-B crosstalk, T cell costimulation, autoantibody production and Th2-mediated Ig production. The kinetics of upregulation of CD86 upon stimulation, supports its major contribution during the primary phase of an immune response.
Applications Reported
This GL1 antibody has been reported for use in flow cytometric analysis.
Applications Tested
This GL1 antibody has been pre-titrated and tested by flow cytometric analysis of LPS stimulated mouse splenocytes using eFluor NC Flow Cytometry Staining Buffer (cat. 00-3222). This can be used at 5 μl per test. A test is defined as the amount 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 105 to 108 cells/test.
The isotype control eFluor 605NC rat IgG2a (cat. 93-4321) should be used at 5μl/test.
Laser/Filter Recommendation: When using eFluor 605NC, we recommend excitation with the 405nm violet laser with an appropriate filter set, such as the 595LP dichroic mirror with the 605/40 bandpass filter. An acceptable alternative is the 610/20 bandpass filter. For instruments not equipped with a violet laser, the eFluor 605NC is also excited by the 488 nm blue laser and can be used as a PE-Texas Red alternative. Please contact eBioscience Technical Support for more information.
Buffer Recommendation: Comparison of eFluor Nanocrystal conjugated antibody staining in different buffers has demonstrated that optimal performance is seen with the eFluor NC Flow Cytometry Staining Buffer (cat. 00-3222). For a comparison of staining with different buffers, refer to eFluor® Nanocrystals page.
Fixation Recommendation: When fixing samples that have been stained with nanocrystal reagents, we recommend keeping the total volume at approximately 200 μl for fixation and the exposure time 30-60 minutes to preserve the optimal fluorescent signal from the nanocrystal reagent.
Hathcock, K. S., G. Laszlo, et al. (1993). "Identification of an alternative CTLA-4 ligand costimulatory for T cell activation [see comments]." Science 262(5135): 905-7. Freeman, G. J., F. Borriello, et al. (1993). "Murine B7-2, an alternative CTLA4 counter-receptor that costimulates T cell proliferation and interleukin 2 production." J Exp Med 178(6): 2185-92. Inaba, K., M. Witmer-Pack, et al. (1994). "The tissue distribution of the B7-2 costimulator in mice: abundant expression on dendritic cells in situ and during maturation in vitro." J Exp Med 180(5): 1849-60. Hathcock, K. S., G. Laszlo, et al. (1994). "Comparative analysis of B7-1 and B7-2 costimulatory ligands: expression and function." J Exp Med 180(2): 631-40