Invitrogen™ CD86 (B7-2) Monoclonal Antibody (GL1), Super Bright™ 702, eBioscience™

Catalog No.	Quantity
67-086-282	100 μg

Catalog No. 67-086-282 Supplier Invitrogen™ Supplier No. 67086282

Description

Rat Monoclonal Antibody

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: This GL1 antibody has been reported for use in flow cytometric analysis. Applications Tested: This GL1 antibody has been tested by flow cytometric analysis of stimulated mouse splenocytes. This may be used at less than or equal to 0.25 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. Super Bright 702 is a tandem dye that can be...

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.

Specifications

• Antigen: CD86 (B7-2)
• Applications: Flow Cytometry
• Classification: Monoclonal
• Clone: GL1
• Concentration: 0.2 mg/mL
• Conjugate: Super Bright 702
• Formulation: PBS with BSA and 0.09% sodium azide; pH 7.2
• Gene: CD86
• Gene Accession No.: P42082
• Gene Alias: activation B7-2 antigen; B lymphocyte activation antigen B72; B7; B7.2; B70; B7-2; B-lymphocyte activation antigen B7-2; BU63; CD28 antigen ligand 2; Cd28l2; CD28LG2; CD86; CD86 antigen; CD86 antigen (CD28 antigen ligand 2, B7-2 antigen); CD86 antigen precursor; CD86 molecule; CLS1; CTLA-4 counter-receptor B7.2; early T cell costimulatory molecule-1; early T-cell costimulatory molecule 1; Early T-cell co-stimulatory molecule 1; ETC-1; FUN-1; LAB72; Ly58; Ly-58; MB7; MB7-2; membrane glycoprotein; MGC34413; T-lymphocyte activation antigen CD86; TS/A-2
• Gene Symbols: CD86
• Host Species: Rat
• Purification Method: Affinity chromatography
• Quantity: 100 μg
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 12524
• Target Species: Mouse
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG2a κ

Frequently Asked Questions (FAQs)

Can I use the OneComp and UltraComp eBeads microspheres with Super Bright-conjugated antibodies?

UltraComp eBeads microspheres (Cat. No. 01-2222) are recommended for use with Super Bright dyes.
Note: Super Bright Staining Buffer (Cat. No. SB-4400) is not compatible with UltraComp eBeads microspheres (Cat. No. 01-2222-41, 00-2222-42). If using UltraComp eBeads microspheres as a compensation tool, solely use Flow Cytometry Stain Buffer (Cat. No. 00-4222-26, 00-4222-57) for any antibody dilutions.

In some experiments, we have observed that compensation values for Super Bright 780- and Brilliant Violet 785- or Brilliant Violet 786-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads, both with Super Bright 780 and Brilliant Violet 786. We have not tested Brilliant Violet 785 with the AbC beads.

Can I prepare an antibody cocktail containing Super Bright Staining Buffer and Super Bright-conjugated antibodies ahead of time?

We recommend that the antibody cocktails containing Super Bright-conjugated antibodies and Super Bright Staining Buffer are prepared fresh prior to staining. Discard any unused portions. We do not recommend overnight storage of prepared cocktails.

Can I fix my cells after staining with Super Bright-conjugated antibodies? How long can fixed cells be stored prior to analysis?

Samples that have been stained with antibodies conjugated to Super Bright dyes may be stored for up to three days, at 2-8°C, in the dark, using either IC Fixation Buffer (Cat. No. 00-8222) or 1-step Fix/Lyse Buffer (Cat. No. 00-5333) with no significant effect on brightness or compensation.

Are the Super Bright dyes sensitive to methanol fixation?

Super Bright dyes are stable in methanol-based fixation buffers.

Can Super Bright-conjugated antibodies be used in combination with intracellular (IC Fixation/Permeabilization) or intranuclear (Foxp3 fixation/permeabilization) staining?

Yes, Super Bright-conjugated antibodies are stable in formaldehyde-based fixation buffers and permeabilization buffers, such as the IC Fixation and Permeabilization Buffer Set (Cat. No. 88-8824) and the Foxp3/Transcription Factor Staining Buffer Set (Cat. No. 00-5523).

Which laser do I use to detect Super Bright-conjugated antibodies?

The violet laser (405 nm) should be used to excite Super Bright-conjugated antibodies.

Can the Super Bright Staining Buffer be used with other polymer dyes?

Yes, the Super Bright Staining Buffer (Cat. No. SB-4400) is compatible with other polymer dyes (i.e., Brilliant Violet dyes) and is useful for minimizing any non-specific polymer interactions when two or more of these dyes are used in combination.

Can Super Bright-conjugated antibodies be used in combination with other polymer dyes? What buffer should I use when using more than one Super Bright or polymer dye?

When using two or more Super Bright dyes, or when using Super Bright dyes in combination with other polymer dyes (i.e., Brilliant Violet dyes), we recommend using the Super Bright Staining Buffer (Cat. No. SB-4400) to minimize any non-specific polymer interactions between these fluorochromes. For specific instructions for use, please refer to the product technical data sheet.

Are there recommended tubes for staining with Super Bright-conjugated antibodies?

We recommend using polystyrene tubes (for example, FACS tubes) for staining with your Super Bright-conjugated antibodies. If using polypropylene (for example, Eppendorf tubes), protecting from light is critical.

Is there a specific buffer I should use when staining with Super Bright-conjugated antibodies?

No special buffer is required when using a single Super Bright-conjugated antibody in a panel.
When using more than one Super Bright dye, or when using Super Bright dyes in combination with other polymer dyes (i.e., Brilliant Violet dyes), we recommend using the Super Bright Staining Buffer (Cat. No. SB-4400) to minimize any non-specific polymer interactions between these fluorochromes. For specific instructions for use, please refer to the product technical data sheet.

How does the protocol for staining with Super Bright-conjugated antibodies differ from staining with conventional organic fluorochrome and eVolve conjugated antibodies?

Antibodies conjugated to Super Bright dyes can be used similarly to traditional fluorochromes. If multiple Super Bright dyes are used in combination with each other or in combination with other polymer dyes (i.e., Brilliant Violet dyes), then the use of Super Bright Staining Buffer (Cat. No. SB-4400) is recommended to minimize any non-specific polymer interactions between these fluorochromes. For specific instructions for use, please refer to the product technical data sheet.

Can the Super Bright dyes be used for non-flow applications?

Super Bright dyes have not been tested for applications other than flow cytometry.

What is the difference between the Super Bright and eVolve dyes?

Super Bright dyes are fluorochromes based on polymer technology and are excited by the violet laser. In contrast, eVolve dyes are based on Qdot nanocrystal technology and are maximally excited by the UV laser, although they may also be excited by violet, blue, yellow-green, and, depending on the eVolve, the red laser lines. The general shape of the emission spectra for Super Bright dyes will be more similar to traditional fluorochromes, while eVolve dyes have extremely narrow emission spectra. Please contact Tech Support (techsupport@thermofisher.com) for more information.

What are the Super Bright dyes?

eBioscience Super Bright dyes are a series of patent-pending fluorochromes that are based upon a fluorescent polymer and its tandems. Super Bright dyes have been developed for use in flow cytometry and can be excited by the violet laser (405 nm). The peak emission of each Super Bright dye is indicated by the number in the product description, e.g., "Super Bright 600" has a peak emission of 600 nm.

Are the Super Bright Dyes photo-labile?

As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.