Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Companion Products
The 29F.1A12 monoclonal antibody specifically recognizes CD279 which is also known as Programmed Death-1 (PD-1). CD279 (PD-1) is a ~55-kDa type I transmembrane glycoprotein that is encoded by Pdcd1 (Programmed cell death 1) which belongs to the CD28/CTLA-4 family of immunoreceptors within the Ig superfamily. CD279 (PD-1) is comprised of an extracellular region with an IgV-like domain, a transmembrane sequence, and an intracellular region with an immunoreceptor tyrosine-based inhibitory motif (ITIM) and an immunoreceptor tyrosine-based switch motif (ITSM) that are associated with suppressive immunoregulatory functions. CD279 (PD-1) is variably expressed on some thymocyte subsets and developing B lymphocytes at the pro-B-cell stage. It is also inducibly expressed on activated myeloid cells, B cells, and T cells including exhausted T cells found in mice during chronic viral infections or cancer. Although this co-inhibitory receptor plays roles in mediating immunological tolerance and preventing autoimmune responses it can also inhibit protective immune responses against microbial infections and cancer. CD273 (also known as PD-L2 or B7-DC) and CD274 (PD-L1 or B7-H1) are members of the B7 family within the Ig superfamily. These molecules serve as ligands for CD279 (PD-1) and are variably expressed on lymphoid and nonlymphoid cell types including antigen-presenting cells and tumor cells. The 29F.1A12 antibody can reportedly block the binding of these ligands as well as other mouse PD-1-specific antibodies including clones J43, G4, and RMP1-14. Antibody-mediated inhibition of the interaction between PD-1 and its ligands can serve as an immune checkpoint blockade that can augment T-cell responses against tumor cells.
Development References (4)
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Liang SC, Latchman YE, Buhlmann JE, et al. Regulation of PD-1, PD-L1, and PD-L2 expression during normal and autoimmune responses.. Eur J Immunol. 2003; 33(10):2706-16. (Immunogen: Flow cytometry, Immunofluorescence, Immunohistochemistry). View Reference
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Lázár-Molnár E, Gácser A, Freeman GJ, Almo SC, Nathenson SG, Nosanchuk JD. The PD-1/PD-L costimulatory pathway critically affects host resistance to the pathogenic fungus Histoplasma capsulatum.. Proc Natl Acad Sci U S A. 2008; 105(7):2658-63. (Clone-specific: In vivo exacerbation). View Reference
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Polesso F, Munks MW, Rott KH, Smart S, Hill AB, Moran AE. PD-1-specific "Blocking" antibodies that deplete PD-1+ T cells present an inconvenient variable in preclinical immunotherapy experiments.. Eur J Immunol. 2021; 51(6):1473-1481. (Clone-specific). View Reference
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Puzey MS, Craig CJ. Hydrometrocolpos--a case report.. S Afr Med J. 1992; 81(6):336-7. (Biology). View Reference
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