CD27 Monoclonal Antibody (O323), PerCP-eFluor™ 710, eBioscience™, Invitrogen™

Description

• Description: The O323 monoclonal antibody reacts with human CD27, a lymphocyte-specific member of the TNFR superfamily
• CD27 is expressed by a subset of thymocytes and virtually all mature T cells and is upregulated upon T-cell stimulation
• CD27 binds to CD70, and through this interaction, plays an important role in T cell-B cell interaction
• Applications Reported: This O323 antibody has been reported for use in flow cytometric analysis
• Applications Tested: This O323 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells
• This can be used at 5 μL (0.06 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
• PerCP-eFluor™ 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5
• We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative
• Please make sure that your instrument is capable of detecting this fluorochrome
• Fixation: Samples can be stored in IC Fixation Buffer (cat
• 00-8222) (100 μL cell sample + 100 μL IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat
• 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation
• CD27 is a 50 kDa member of the tumor necrosis factor (TNF) receptor superfamily that includes CD40 and CD30
• The TNF superfamily members are known for the regulation of cell proliferation and death
• In contrast to the expression of other TNFR/TNF family members, expression of CD27 and its ligand CD70 is predominantly confined to lymphocytes
• High expression levels of CD27 appear to be dependent on proper ligation of antigen receptors
• CD70 expression requires additional co-stimulatory and/or pro-inflammatory signals
• CD27 is expressed as a disulfide-linked homodimer on mature thymocytes, peripheral blood T cells and a subpopulation of B cells
• Activation of T cells via TCR-CD3 complex results in upregulation of CD27 expression on the plasma membrane as well as in the release of its soluble 28-32 kDa form, sCD27, detected in the plasma, urine or spinal fluid
• Soluble CD27 is an important prognostic marker of acute and chronic B cell malignancies
• RgpA, a cystein proteinase, although activating T cells through the protease-activated receptors (PARs), degradates CD27 and counteracts T cell activation mediated by CD27 and its ligand CD70
• CD27-binding protein (SIVA), a proapoptotic protein, can bind to this receptor and is thought to play an important role in the apoptosis induced by this receptor
• Diseases associated with CD27 dysfunction include Lymphoproliferative Syndrome 2 and Autosomal Recessive Lymphoproliferative Syndrome.