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Invitrogen
CD28 Monoclonal Antibody (CD28.2), NovaFluor™ Yellow 660, eBioscience™
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FIGURE: 1 / 3
CD28 Antibody (H008T03Y04-A) in Flow
Normal human peripheral blood cells were unstained (left) or stained with CD28 Monoclonal Antibody, NovaFluor Yellow 660 (right). All cells were co-stained with CD19 Monoclonal Antibody, eFluor 450 (Product # 48-0199-42). Total viable cells were used for analysis, as determined by LIVE/DEAD Blue (Product # L34962). Data was acquired in the YG4 channel on a 5-laser Cytek Aurora for the NovaFluor Yellow 660 conjugate. Cells in the lymphocyte gate were used for analysis.
Product Details
H008T03Y04-A
Product Specifications
Species Reactivity
Human
Host/Isotype
Mouse
/ IgG1, kappa
Class
Monoclonal
Type
Antibody
Clone
CD28.2
Conjugate
Excitation/Emission Max
550/664 nm
View spectra
Form
Liquid
Concentration
5 µL/Test
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.2
Contains
0.09% sodium azide
Storage conditions
4°C, store in dark, DO NOT FREEZE!
RRID
AB_3098114 (AB_2896630)
Product Specific Information
Description
The CD28.2 monoclonal antibody reacts with the human CD28 molecule, a 44 kDa homodimer expressed by thymocytes, mature T cells and plasma cells. CD28 is a ligand for CD80 (B7-1) and CD86 (B7-2) and is a potent co-stimulator of T cells. Signaling through CD28 augments IL-2 and IL-2 receptor expression as well as cytotoxicity of CD3-activated T cells.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
This CD28.2 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.8 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 (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: 550 nm; Emission: 664 nm; Laser: 561 nm (Yellow) Laser
Target Information
CD28 antigen is a 44 kDa disulfide linked homodimeric T cell specific surface glycoprotein. CD28 is a cell adhesion molecule of the immunoglobulin superfamily which is constitutively expressed on most peripheral blood T lymphocytes. Moreover, CD28 is the critical T cell costimulatory receptor that provides the cell the important second activation signal by binding CD80 and CD86 which are expressed by antigen presenting cells. In addition to its co-stimulation role, CD28 functions by preventing T cells from entering an anergic-hyporesponsive state or from undergoing premature apoptotic cell death. In murine peripheral lymphoid organs and in the blood, all CD4+ and CD8+ T cells express CD28. In the thymus, CD28 expression is highest on immature CD3-, CD8+ and CD4+8+ cells, and on CD4-8- cells that express alpha/beta and gamma/delta TCR. The level of CD28 on mature CD4+ and CD8+ alpha/beta TCR+ thymocytes is two- to fourfold lower than on the immature cells. Diseases associated with CD28 dysfunction include mycosis fungiodes and Sezary's Disease.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
How to use the Panel Builder
Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.
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Bioinformatics
Protein Aliases: CD28; CD28 antigen; MGC138290; sCD28; soluble CD28; T-cell-specific surface glycoprotein CD28; T44; TP44
Gene Aliases: CD28; Tp44
UniProt ID: (Human) P10747
Entrez Gene ID: (Human) 940
positive regulation of inflammatory response to antigenic stimulus
humoral immune response
cell surface receptor signaling pathway
positive regulation of gene expression
negative regulation of gene expression
regulation of phosphatidylinositol 3-kinase signaling
positive regulation of phosphatidylinositol 3-kinase signaling
T cell costimulation
positive regulation of interleukin-10 production
positive regulation of interleukin-4 production
cytokine biosynthetic process
positive regulation of T cell proliferation
negative thymic T cell selection
regulatory T cell differentiation
positive regulation of viral genome replication
positive regulation of interleukin-2 biosynthetic process
regulation of regulatory T cell differentiation
positive regulation of translation
positive regulation of mitotic nuclear division
positive regulation of transcription from RNA polymerase II promoter
positive regulation of alpha-beta T cell proliferation
phosphatidylinositol phosphorylation
phosphatidylinositol-mediated signaling
positive regulation of isotype switching to IgG isotypes
regulation of defense response to virus by virus
T cell receptor signaling pathway
positive regulation of protein kinase B signaling
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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