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CD86 (B7-2) Monoclonal Antibody (IT2.2), NovaFluor™ Blue 760, eBioscience™
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FIGURE: 1 / 2
CD86 (B7-2) Antibody (H021T03B11-A) in Flow
Normal human peripheral blood cells were unstained (left) or stained with CD86 (B7-2) Monoclonal Antibody, NovaFluor Blue 760 (right). All cells were co-stained with CD14 Monoclonal Antibody, eFluor 450 (Product # 48-0149-42). Total viable cells in the monocyte 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 autofluorescence extraction.
Product Details
H021T03B11-A
Product Specifications
Species Reactivity
Human
Host/Isotype
Mouse
/ IgG2b, kappa
Class
Monoclonal
Type
Antibody
Clone
IT2.2
Conjugate
Excitation/Emission Max
490/764 nm
View spectra
Form
Liquid
Concentration
0.6 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 IT2.2 monoclonal antibody reacts with human CD86, an ~80 kDa surface receptor also known as B7-2.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
This IT2.2 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.6 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: 493 nm; Emission: 764 nm; Laser: 488 nm (Blue) Laser
Target Information
CD86, along with CD80, is a member of the B7 family of costimulatory molecules and plays a crucial role in T cell activation and immune response regulation. CD86 is expressed at low levels on B cells, macrophages, and dendritic cells, and its expression is upregulated on B cells through various stimuli, including the BCR complex, CD40, and certain cytokine receptors. As a type I membrane protein and member of the immunoglobulin superfamily, CD86 serves as a ligand for the T cell surface proteins CD28 and CTLA-4 (CD152). The interaction between CD86 and CD28 provides a costimulatory signal essential for T cell activation during antigen presentation, while binding with CTLA-4 negatively regulates T cell activation, diminishing the immune response. This interaction is critical for T-B cell crosstalk, T cell costimulation, autoantibody production, and Th2-mediated Ig production. The kinetics of CD86 upregulation upon stimulation suggest its significant contribution during the primary phase of an immune response. CD86 and CD80 have distinct roles in T helper cell differentiation, and insufficient co-stimulation involving these molecules can induce tolerance. Alternative splicing of CD86 results in two transcript variants encoding different isoforms, with additional variants described but not fully sequenced. Dysfunction in CD86 is associated with diseases such as gallbladder squamous cell carcinoma and myocarditis.
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: Activation B7-2 antigen; B lymphocyte activation antigen B72; B-lymphocyte activation antigen B7-2; B70; BU63; CD28 antigen ligand 2; CD86; CD86 antigen (CD28 antigen ligand 2, B7-2 antigen); CD86 v6; CTLA-4 counter-receptor B7.2; Early T-cell co-stimulatory molecule 1; FUN-1; MGC34413; T-lymphocyte activation antigen CD86; unnamed protein product
Gene Aliases: B7-2; B7.2; B70; CD28LG2; CD86; LAB72
UniProt ID: (Human) P42081
Entrez Gene ID: (Human) 942
adaptive immune response
immune system process
positive regulation of immunoglobulin production
immune response
signal transduction
cell surface receptor signaling pathway
positive regulation of cell proliferation
T cell costimulation
positive regulation of interleukin-2 production
positive regulation of interleukin-4 production
positive regulation of lymphotoxin A production
positive regulation of T cell proliferation
T cell activation
B cell activation
negative regulation of T cell proliferation
positive regulation of T-helper 2 cell differentiation
positive regulation of transcription, DNA-templated
lymphocyte activation
viral entry into host cell
cellular response to lipopolysaccharide
positive regulation of NIK/NF-kappaB signaling
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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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