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60712
CD86/B7-2 (GL-1) Rat mAb (PE Conjugate)
Antibody Conjugates

CD86/B7-2 (GL-1) Rat mAb (PE Conjugate) #60712

APPLICATIONS

REACTIVITY SENSITIVITY MW (kDa) Isotype
M Endogenous Rat IgG2a, kappa
Flow Cytometry

Flow cytometric analysis of live mouse splenocytes, untreated (left column) or treated with LPS #14011 (500 ng/ml, 3 d; right column), using CD86/B7-2 (GL-1) Rat mAb (PE Conjugate) (top row) or concentration-matched Rat Isotype Control (PE Conjugate) (bottom row), and co-stained with CD19 (ID3) Rat mAb (APC Conjugate) #39831.

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Flow Cytometry

A. Solutions and Reagents

NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.

  1. 20X Phosphate Buffered Saline (PBS): (#9808) To prepare 1 L 1X PBS: add 50 ml 20X PBS to 950 ml dH2O, mix.
  2. Incubation Buffer: Dissolve 0.5 g Bovine Serum Albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.

B. Immunostaining

  1. Count cells using a hemacytometer or alternative method.
  2. Aliquot 0.5-1x106 cells into each assay tube (by volume).
  3. Add 2-3 ml Incubation Buffer to each tube and rinse by centrifugation. Repeat.
  4. Resuspend cells in 100 µl of diluted primary antibody (prepared in incubation buffer at the recommended dilution).
  5. Incubate for 1 hr at room temperature.
  6. Wash by centrifugation in 2-3 ml incubation buffer.
  7. Resuspend cells in the appropriate volume of PBS and analyze on flow cytometer.

posted June 2017

Protocol Id: 1504

For optimal flow cytometry results, we recommend 0.125 μg of antibody per test.

Application Dilutions
Flow Cytometry 1:160
Storage:

Supplied in 10 mM NaH2PO4, 150 mM NaCl, 0.09% NaN3, 0.1% gelatin, pH 7.2. This product is stable for 6 months when stored at 4ºC. Do not aliquot the antibody. Protect from light. Do not freeze.

CD86/B7-2 (GL-1) Rat mAb (PE Conjugate) recognizes endogenous levels of total CD86/B7-2 protein. This antibody detects an epitope within the extracellular domain.

Species Reactivity:

Mouse

This monoclonal antibody was purified from tissue culture supernatant via affinity chromatography. The purified antibody was conjugated under optimal conditions, with unreacted dye removed from the preparation.

This Cell Signaling Technology antibody is conjugated to PE and tested in-house for direct flow cytometric analysis in mouse cells.

CD80 (B7-1, BB1) and CD86 (B7-2, B70) are members of the B7 family of cell surface ligands that regulate T cell activation and immune responses. CD80 is expressed on activated antigen presenting cells, including dendritic cells, B cells, monocytes, and macrophages. CD86 is expressed on resting monocytes, dendritic cells, activated B lymphocytes, and can be further upregulated in the presence of inflammation (1-3). CD80 and CD86 are ligands for CD28, which functions as a T cell costimulatory receptor. Interaction of CD28 with CD80 or CD86 provides the second signal required for naïve T cell activation, T cell proliferation, and acquisition of effector functions (3-7). Alternatively, CD80 and CD86 also act as ligands to CTLA-4, which results in the downregulation of T cell activity (3,7-9).

The GL-1 antibody is used as a marker for CD86 expression on B cells, macrophages, and dendritic cells.

  1. Freeman, G.J. et al. (1989) J Immunol 143, 2714-22.
  2. Azuma, M. et al. (1993) Nature 366, 76-9.
  3. Adams, A.B. et al. (2016) J Immunol 197, 2045-50.
  4. Linsley, P.S. et al. (1990) Proc Natl Acad Sci U S A 87, 5031-5.
  5. Gimmi, C.D. et al. (1991) Proc Natl Acad Sci U S A 88, 6575-9.
  6. Harding, F.A. et al. (1992) Nature 356, 607-9.
  7. Collins, A.V. et al. (2002) Immunity 17, 201-10.
  8. Linsley, P.S. et al. (1991) J Exp Med 174, 561-9.
  9. Krummel, M.F. and Allison, J.P. (1995) J Exp Med 182, 459-65.
Entrez-Gene Id
12524
Swiss-Prot Acc.
P42082
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

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To Purchase # 60712S

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Product # Size Price
60712S
100 µg  (10 western blots) $ 189.0