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Invitrogen
CD27 Monoclonal Antibody (O323), NovaFluor™ Violet 690, eBioscience™
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FIGURE: 1 / 2
CD27 Antibody (H012T03V01-A) in Flow
Normal human peripheral blood cells were unstained (left) or stained with CD27 Monoclonal Antibody, NovaFluor Violet 690 (right). All cells were co-stained with CD19 Monoclonal Antibody, eFluor 450 (Product # 48-0199-42). Total viable cells in the lymphocyte gate were used for analysis, as determined by LIVE/DEAD Blue (Product # L34962). Data was acquired on a 5-laser Cytek Aurora and unmixed with multiple autofluorescence signature extraction.
Product Details
H012T03V01-A
Product Specifications
Species Reactivity
Human
Host/Isotype
Mouse
/ IgG1, kappa
Class
Monoclonal
Type
Antibody
Clone
O323
Conjugate
Excitation/Emission Max
408/691 nm
View spectra
Form
Liquid
Concentration
0.4 µg/Test
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.2, with BSA
Contains
0.09% sodium azide
Storage conditions
4°C, store in dark, DO NOT FREEZE!
RRID
Product Specific Information
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.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
The 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 (0.4 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.
Master mixes
• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.
• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.
Whole Blood compatibility
• When utilizing whole blood (as opposed to density-gradient-purified PBMC), we recommend lysing red blood cells in bulk prior to staining with NovaFluor conjugates.
• See the Bulk Lysis of Human Whole Blood protocol here.
• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.
Viability dye compatibility
• NovaFluor dyes are not compatible with DNA intercalating viability dyes.
• Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.
CellBlox Plus Blocking Buffer
• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (for 25 µg or 25 test products, Cat. No. C001T02F01; for 100 µg or 100 test, Cat. No. C001T03F01), essential for optimal staining.
• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.
• Add 5 µL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.
• CellBlox Plus Blocking Buffer is compatible with either Super Bright Complete Blocking Buffer or Brilliant Stain Buffer and can be used in antibody cocktails/master mixes with those reagents.
• For single-color controls, use 5 µL of CellBlox Plus Blocking Buffer per 100 µL of cell sample (10^3 to 10^8 cells).
NovaFluor conjugates are based on Phiton technology utilizing novel fluorophore-containing nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more
Excitation: 408 nm; Emission: 691 nm; Laser: 405 nm (Violet) Laser
Target Information
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.
How to use the Panel Builder
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Bioinformatics
Protein Aliases: CD27; CD27 antigen; CD27L receptor; LPFS2; sCD27; soluble CD27; T cell activation antigen S152; T-cell activation antigen CD27; T14; TNFSF7; 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
inflammatory response
immune response
cell surface receptor signaling pathway
brain development
release of cytoplasmic sequestered NF-kappaB
extrinsic apoptotic signaling pathway via death domain receptors
immunoglobulin mediated immune response
response to lipopolysaccharide
tumor necrosis factor-mediated signaling pathway
regulation of cell proliferation
negative regulation of apoptotic process
negative regulation of cysteine-type endopeptidase activity involved in apoptotic process
positive regulation of MAPK cascade
response to ethanol
positive regulation of B cell differentiation
positive regulation of JNK cascade
negative regulation of T cell apoptotic process
extrinsic apoptotic signaling pathway
Performance Guarantee
If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*
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