Invitrogen

CD86 (B7-2) Monoclonal Antibody (GL1), PE-Cyanine5, eBioscience™

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CD86 (B7-2) Antibody — Fig. 8 4-TMPs activate DCs regardless of TCR engagement. a Representative confocal images of Fluo-3-loaded DCs placed on a 4-TMP-deposited lipid bilayer. b Fluo-3-loaded DCs from wild type or TLR4 -/- mice were stimulated with 4-TMPs purified from CD4 + T cells in the presence or absence of SEB. Calcium changes were analysed by flow cytometry for 300 s. Data are representative of at least three separate experiments. c Surface protein expression on DCs induced by 4-TMPs. DCs (5 x 10 6 ) from wild-type mice were treated with 4-TMPs obtained from CD4 + T cells (0.05-1 x 10 8 ) at the indicated ratio (DC:T = 1:1 to 1:20). After 24 h, the expression of CD86 was analysed by flow cytometry and expressed as the mean fluorescence intensity (MFI). Data represent the mean of three experiments +- SEM. * P < 0.01 vs. 1:0. d DCs (5 x 10 6 ) from wild type or TLR4 -/- mice were treated with 4-TMPs (5 x 10 7 ). After 24 h, the expression of CD86, CD40, and CD80 was determined as described in c . In some cases, FcgammaR was blocked 30 min before 4-TMP treatment. * P < 0.01 vs. PBS-treated group

CD86 (B7-2) Antibody in Flow Cytometry (Flow) — Staining of 3-day LPS activated BALB/c splenocytes with 0.03 µg of Rat IgG2a Isotype Control PE-Cyanine5 (Product # 15-4321-80) (open histogram) or Anti-Mouse CD86 (B7-2) PE-Cyanine5 (filled histogram).Total viable cells were used for analysis.

Product Details

Tested Applications

Dilution

Flow Cytometry (Flow)

0.06 µg/test

Published Applications

Flow Cytometry (Flow)

See 27 publications below

Product Specifications

Species Reactivity

Mouse

Published species

Mouse

Host / Isotype

Rat / IgG2a, kappa

Recommended Isotype Control

Rat IgG2a kappa Isotype Control (eBR2a), PE-Cyanine5, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

GL1

Conjugate

PE-Cyanine5

Excitation/Emission Profile

View spectra

Form

Liquid

Concentration

0.2 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2, with 0.1% gelatin

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

RRID

AB_468778

Product Specific Information

Description: The GL1 monoclonal antibody reacts with mouse CD86, an ~80 kDa surface receptor also known as B7-2. CD86 and CD80 are members of the B7 family of costimulatory molecules. CD86 is expressed at low level on B cells, macrophages, and dendritic cells and is upregulated on B cells through a variety of surface stimuli including the BCR complex, CD40 and some cytokine receptors. CD86 is also expressed by activated mouse T cells and thioglycolate-elicited peritoneal cells. In addition to CD80 (B7-1), CD86 is a counter-receptor for the T cell surface molecules CD28 and CD152 (CTLA-4). This interaction plays a critical role in T-B crosstalk, T cell costimulation, autoantibody production and Th2-mediated Ig production. The kinetics of upregulation of CD86 upon stimulation, supports its major contribution during the primary phase of an immune response.

Applications Reported: The GL1 antibody has been reported for use in flow cytometric analysis.

Applications Tested: The GL1 antibody has been tested by flow cytometric analysis of resting and activated mouse splenocytes. This can be used at less than or equal to 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

Light sensitivity: This tandem dye is sensitive photo-induced oxidation. Please protect this vial and stained samples from light.

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. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.

Excitation: 488-561 nm; Emission: 667 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

CD86 is one of two ligands (the other CD80) for CTLA4 and CD28. CD86 acts as costimulatory molecule in eliciting T-cell help during antigen presentation. Antigen presentation in the absence of sufficient co-stimulation involving CD86/CD80 can induce tolerance. CD80 appears to play a role distinct from CD80 in T helper cell differentiation. CD86 is a type I membrane protein that is a member of the immunoglobulin superfamily. The CD86 protein is expressed by antigen-presenting cells, and it is the ligand for two proteins at the cell surface of T cells, CD28 antigen and cytotoxic T-lymphocyte-associated protein 4. Binding of CD86 with CD28 antigen is a costimulatory signal for activation of the T-cell. Binding of CD86 with cytotoxic T-lymphocyte-associated protein 4 negatively regulates T-cell activation and diminishes the immune response. Alternative splicing results in two transcript variants encoding different isoforms of CD86. Additional transcript variants have been described for CD86, but their full-length sequences have not been determined. Diseases associated with CD86 dysfunction include gallbladder squamous cell carcinoma and myocarditis.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: Activation B7-2 antigen; B-lymphocyte activation antigen B7-2; B-lymphocyte antigen B7-2; B7-2; B7-2 antigen; B7.2; CD28 antigen ligand 2; CD28LG2; CD86; early T cell costimulatory molecule-1; Early T-cell costimulatory molecule 1; ETC-1; LAB72; MGC34413; T-lymphocyte activation antigen CD86

Gene Aliases: B7; B7-2; B7.2; B70; Cd28l2; Cd86; CLS1; ETC-1; Ly-58; Ly58; MB7; MB7-2; TS/A-2

UniProt ID: (Mouse) P42082

Entrez Gene ID: (Mouse) 12524

Function(s)

toll-like receptor signaling pathway immune system process negative regulation of T cell anergy positive regulation of T cell proliferation positive regulation of activated T cell proliferation B cell activation defense response to virus cellular response to lipopolysaccharide

Process(es)

receptor binding

CD86 (B7-2) Monoclonal Antibody (GL1), PE-Cyanine5, eBioscience™

Product Details

Product Specific Information

Target Information

Bioinformatics

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