CD279 (PD-1) Monoclonal Antibody (eBioJ105 (J105)), Brilliant Ultra Violet™ 615, eBioscience™

Product Specific Information

Description
The J105 monoclonal antibody reacts with the human PD-1 (programmed death-1), a 55 kDa member of the CD28 immunoglobulin superfamily. PD-1 contains the immunoreceptor tyrosine-based inhibitory motif (ITIM) and plays a key role in peripheral tolerance and autoimmune disease. PD-1 is expressed predominantly on activated T and B lymphocytes. Two novel members of the B7 family have been identified as the PD-1 ligands, PD-L1 (B7-H1) and PD-L2 (B7-DC). Evidence reported to date suggests overlapping functions for these two PD-1 ligands and their constitutive expression on some normal tissues and upregulation on activated antigen-presenting cells. Costaining experiments suggest that eBioJ105 recognizes a different epitope than MIH4 (Product # 11-9969).

Applications Tested
This eBioJ105 (J105) antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may 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.

Blocking Buffers
When using two or more Super Bright, Brilliant Violet™, Brilliant Ultra Violet™, or other polymer dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) or Brilliant Stain Buffer (Product # 00-4409-75) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer or Brilliant Stain 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: 350 nm; Emission: 615 nm; Laser: Ultraviolet Laser.

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Target Information

Cell-mediated immune responses are initiated by T lymphocytes that are themselves stimulated by cognate peptides bound to MHC molecules on antig en-presenting cells (APC). T-cell activation is generally self-limited as activated T cells express receptors such as PD-1 (also known as PDCD-1) that mediate inhibitory signals from the APC. PD-1 can bind two different but related ligands, PDL-1 and PDL-2. Upon binding to either of these ligands, signals generated by PD-1 inhibit the activation of the immune response in the absence of "danger signals" such as LPS or other molecules associated with bacteria or other pathogens. Evidence for this is seen in PD1-null mice who exhibit hyperactivated immune systems and autoimmune diseases. Despite its predicted molecular weight, PD-1 often migrates at higher molecular weight in SDS-PAGE.