Description
The PO3 monoclonal antibody specifically binds to the B7-2 (CD86) costimulatory molecule, which is expressed on a broad spectrum of leukocytes, including B lymphocytes, T lymphocytes, macrophages, dendritic cells, and Langerhans cells, plus astrocytes. CD86 is expressed at low levels by freshly explanted peripheral B and T cells, and its expression is substantially increased by a variety of T and B cell-specific stimuli with a peak expression after 18-42 hours of culture. In contrast to most naive CD4+ T cells, memory CD4+ T cells express B7-2, both at the mRNA and protein levels. CD86, a ligand for CD28 and CTLA-4, is one of the accessory molecules that plays an important role in T cell-B cell costimulatory interactions. It has been shown to be involved in immunoglobulin class switching and triggering of mouse NK cell-mediated cytotoxicity. CD80 (B7-1) is an alternate ligand for CD28 and CTLA-4. PO3 antibody blocks the in vitro stimulation of T-cell proliferation of T-cell proliferation by soluble anti-CD3e antibody (mAb 145-2C11, Cat. No. 553057) in the presence of CD86-expressing accessory cells. In vivo administration of PO3 mAb can inhibit much of the autoantibody production in (NZB x NZW) F1 mice; and in combination with an anti-CD80 mAb, it can prevent the development and progression of mouse systemic lupus erythematosus-like autoimmune disease.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794 or 566349). When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values. For optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent staining prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon BB700 under optimal conditions that minimize unconjugated dye and antibody.
1. This antibody was developed for use in flow cytometry.
2. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
3. Researchers should determine the optimal concentration of this reagent for their individual applications.
4. An isotype control should be used at the same concentration as the antibody of interest.
5. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
8. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
9. BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
10. Cy is a trademark of GE Healthcare.
