CD86 Antibody, anti-mouse, REAfinity™

Name: CD86 Antibody, anti-mouse, REAfinity™
Availability: InStock

CD86 Antibody, anti-mouse, REAfinity™Extended ValidationRecombinant

Clone:

REA1190

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MICS, IF, IHC

Alternative names:

B7-2, B70, CLS1, Cd28l2, ETC-1, Ly-58, MB7-2, TS/A-2

Flow cytometry applications forCD86 Antibody, anti-mouse, REAfinity™

APC
PE
Vio Bright B515
Vio Bright R720

Conjugate:

APC

Recommended dilution:

1:50

Remark:

The antibody is suited for staining of formaldehyde-fixed cells.

Tested:

QC tested

Products used:

130-122-130

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:50)

Description:

Splenocytes of BALB/c mice were separated with the Spleen Dissociation Kit (130-095-926). The positive fraction was stimulated with 100 ng/mL LPS for 24 hours. A mixture of stimulated and unstimulated cells were then stained with CD86 antibodies or with the corresponding REA Control antibodies (left image), as well as with CD11c antibodies. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

MICS (MACSima Imaging Cyclic Staining) applications forCD86 Antibody, anti-mouse, REAfinity™

APC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

mouse

Products used:

130-122-130

Protocols:

Protocol for MICS experiments

Extended validation for CD86 Antibody, anti-mouse, REAfinity™

Specificity

Epitope competition

In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.

Other clones	Overlap in epitope recognition with REA1190
GL1	++
PO3.1	++
PO3.2	++
A17199A	++
Cells were incubated with an excess of purified unconjugated CD86 (REA1190) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison

Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.

Flow cytometric comparison of different clones for CD86. A mixture of CD11c (+) stimulated with 100ng/ml LPS for 24 hours and CD11c (-) unstimulated Balb/c Splenocytes were stained with CD86 antibodies and with a suitable counterstaining. As a control, CD86 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD86. A mixture of CD11c (+) stimulated with 100ng/ml LPS for 24 hours and CD11c (-) unstimulated Balb/c Splenocytes were stained with CD86 antibodies and with a suitable counterstaining. As a control, CD86 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD86. A mixture of CD11c (+) stimulated with 100ng/ml LPS for 24 hours and CD11c (-) unstimulated Balb/c Splenocytes were stained with CD86 antibodies and with a suitable counterstaining. As a control, CD86 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD86. A mixture of CD11c (+) stimulated with 100ng/ml LPS for 24 hours and CD11c (-) unstimulated Balb/c Splenocytes were stained with CD86 antibodies and with a suitable counterstaining. As a control, CD86 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD86. A mixture of CD11c (+) stimulated with 100ng/ml LPS for 24 hours and CD11c (-) unstimulated Balb/c Splenocytes were stained with CD86 antibodies and with a suitable counterstaining. As a control, CD86 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD86. A mixture of CD11c (+) stimulated with 100ng/ml LPS for 24 hours and CD11c (-) unstimulated Balb/c Splenocytes were stained with CD86 antibodies and with a suitable counterstaining. As a control, CD86 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD86. A mixture of CD11c (+) stimulated with 100ng/ml LPS for 24 hours and CD11c (-) unstimulated Balb/c Splenocytes were stained with CD86 antibodies and with a suitable counterstaining. As a control, CD86 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD86. A mixture of CD11c (+) stimulated with 100ng/ml LPS for 24 hours and CD11c (-) unstimulated Balb/c Splenocytes were stained with CD86 antibodies and with a suitable counterstaining. As a control, CD86 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Fixation data

To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.

No fixation

Post fixation

Comparison of staining pattern on non-fixed and fixed cells using CD86 (REA1190). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Comparison of staining pattern on non-fixed and fixed cells using CD86 (REA1190). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Comparison of staining pattern on non-fixed and fixed cells using CD86 (REA1190). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Comparison of staining pattern on non-fixed and fixed cells using CD86 (REA1190). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for CD86 Antibody, anti-mouse, REAfinity™

Overview

Clone REA1190 recognizes the mouse CD86 antigen, also known as B7-2, an 80 kDa molecule and a member of the immunoglobulin superfamily. Together with CD80 (B7-1) it belongs to the B7 family of costimulatory molecules. CD86 is expressed on activated antigen-presenting cells, including B cells, dendritic cells, and monocytes/macrophages. The interaction of CD86 with its ligands CD28 and CD152 (CTLA- 4) plays a critical role in induction and regulation of immune responses, e.g., cross-talk between T and B cells, T cell costimulation, or immunoglobulin class-switching. Binding of CD86 to CD28 on T cells results in transduction of costimulatory signals for activation or proliferation of T cells, or cytokine production. In contrast, binding of CD86 to CTLA-4 regulates T cell activation and diminishes the immune response.

Additional information: Clone REA1190 displays negligible binding to Fc receptors.

Alternative names

B7-2, B70, CLS1, Cd28l2, ETC-1, Ly-58, MB7-2, TS/A-2

Detailed product information

Technical specifications

Clone	REA1190
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody, human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	mouse
Antigen	CD86
Alternative names of antigen	B7-2, B70, CLS1, Cd28l2, ETC-1, Ly-58, MB7-2, TS/A-2
Molecular mass of antigen [kDa]	32
Distribution of antigen	B cells, dendritic cells, macrophages, monocytes, other
Entrez Gene ID	12524
RRID	AB_2819413, AB_2819414, AB_2921893, AB_2819412

Resources for CD86 Antibody, anti-mouse, REAfinity™

Certificates

Please follow this

link

to search for Certificates of Analysis (CoA) by lot number.

References for CD86 Antibody, anti-mouse, REAfinity™

Publications

Kin, N. W. et al. (2006) CD86 stimulation on a B cell activates the phosphatidylinositol 3-kinase/Akt and phospholipase C gamma 2/protein kinase C alpha beta signaling pathways. J Immunol 176(11): 6727-6735
Nakajima, A. et al. (1997) Requirement of CD28-CD86 co-stimulation in the interaction between antigen-primed T helper type 2 and B cells. Int. Immunol. 9(5): 637-644
Zhu, X. Y. et al. (2005)
Blockade of CD86 signaling facilitates a Tʜ2 bias at the maternal-fetal interface and expands peripheral CD4
⁺
CD25
⁺
regulatory T cells to rescue abortion-prone fetuses.
Biol. Reprod. 72(2): 338-345

Applications

Extended validation