Qdot™ Antibody (Ab) conjugates possess a bright fluorescence emission that makes them well suited for the detection of low-abundance extracellular proteins. Approximately the same size as R-phycoerythrin (R-PE) and compatible with existing organic fluorophore conjugates, Qdot™ Ab conjugates can be excited with any wavelength below their emission maximum, but are best excited by UV or violet light. The narrow, symmetric emission profiles of Qdot Ab conjugates allow for minimal compensation when using a single excitation source, and the very long stoke shifts enable better, more efficient multicolor assays using the 405 nm violet laser. Available in multiple colors for use in flow cytometry, these advantages make Qdot Ab conjugates powerful tools for antibody labeling and staining. Staining: Stain cells in any standard staining buffer, such as phosphate buffered saline (PBS) with 1% bovine serum albumin (BSA). Use 1 L of Qdot antibody conjugate per 1 × 10^6 cells in a 100 µL staining volume (20 nM final concentration of Qdot™ nanocrystals). Qdot™ nanocrystal conjugates may be mixed with other antibodies, but use the diluted conjugates on the day of dilution. Qdot nanocrystal conjugates can be used for surface staining applications with most conventional sample preparation reagents, such as Cal-Lyse™ and FIX & PERM™ reagents, with minimal affect on fluorescence. We have observed that some batches of BD FACS™ Lysing Solution have been found to interfere with Qdot™ nanocrystal fluorescence. Each lot has been tested by flow cytometry using human peripheral blood leukocytes. This testing was performed using 1 L of antibody per 1 × 106 cells in a 100 L staining volume (20 nM final concentration of Qdot nanocrystals). Qdot nanocrystal concentration is assigned based on optical density. The isotype control for this antibody is mouse IgG2a, Cat. No. Q10014.
Instrument setup: Qdot Ab conjugates are excited optimally with UV or 405 nm light, although excitation can be obtained with any wavelength below the emission maximum of a given Qdot nanocrystal, such as with a 488-nm laser. Qdot Ab conjugates can be used on cytometers that do not have UV or violet excitation sources as long as they have appropriate emission filters. Make sure the cytometer has an appropriate emission filter for the Qdot Ab conjugate being used; alternate filters can be used as long as they capture the emission maximum, but filter width impacts spectral overlap corrections. And be sure to check for Qdot Ab conjugate emission in any channel that can capture nanocrystal emission off of other lasers on the cytometer. For Cat. No. Q10065: peak excitation 405 (488) nm/peak emission 605 nm; recommended filter 605/20 nm.
Store reagents at 2-8°C in the dark. Do not freeze. Because Qdot nanocrystals are conjugated to biological materials, some loss of activity may be observed with prolonged storage. When stored as instructed, expires six months from date of receipt unless otherwise indicated on product label. Qdot Ab conjugates are photostable, and do not need to be protected from light. However, if using Qdot Ab conjugates in combination with conventional fluorochrome conjugated antibodies, minimize light exposure during handling, incubation with cells, and prior to analysis. The Qdot Ab conjugates contain cadmium and selenium in an inorganic crystalline form. Dispose of the material in compliance with all applicable local, state, and federal regulations for disposal of these classes of material. For more information on the composition of these materials, consult the Safety Data Sheets (SDSs).
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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: CD27; CD27 antigen; CD27L receptor; sCD27; soluble CD27; T cell activation antigen S152; T-cell activation antigen CD27; T14; Tumor necrosis factor receptor superfamily member 7; tumor necrosis factor receptor superfamily, member 7
Gene Aliases: CD27; S152; S152. LPFS2; T14; TNFRSF7; Tp55
UniProt ID: (Human) P26842
Entrez Gene ID: (Human) 939