CD80 Antibody, anti-human

Name: CD80 Antibody, anti-human
Availability: InStock

CD80 Antibody, anti-humanExtended Validation

Clone:

2D10

Type of antibody:

Primary antibodies

Isotype:

mouse IgG1κ

Applications:

FC

Alternative names:

B7, B7-1, BB1, CD28LG, CD28LG1, LAB7

Flow cytometry applications forCD80 Antibody, anti-human

APC
Biotin
PE
PE-Vio 770

Conjugate:

APC

Recommended dilution:

1:50

Remark:

Cells should be stained prior to fixation, if formaldehyde is used as a fixative.

Tested:

QC tested

Products used:

130-122-972, 130-122-928

Protocols:

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

Description:

CD14⁺ monocytes were isolated with CD14 MicroBeads, human, from peripheral blood mononuclear cells (PBMCs) and cultured in the presence of GM-CSF and interleukin 4 (IL-4) for 6 days into inmature monocyte-derived dendritic cells (Mo-DCs). For generation of mature Mo-DCs, cells were cultured for another 24 hours in the presence of TNF-α, IL-6, and IL-1β. Mo-DCs were then stained with CD80 antibodies or with the corresopnding isotype control antibodies (left peak) and analyzed by flow cytometry using the MACSQuant^® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. 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.

Extended validation for CD80 Antibody, anti-human

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 2D10
REA661	++
QA18A16	++
W17149D	++
L307.4	+
Cells were incubated with an excess of purified unconjugated CD80 (2D10) 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 CD80. Peritoneal macrophages from C57BL/6 mice were stained with CD80 antibodies and with a suitable counterstaining. As a control, CD80 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 CD80. Peritoneal macrophages from C57BL/6 mice were stained with CD80 antibodies and with a suitable counterstaining. As a control, CD80 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 CD80. Peritoneal macrophages from C57BL/6 mice were stained with CD80 antibodies and with a suitable counterstaining. As a control, CD80 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 CD80. Peritoneal macrophages from C57BL/6 mice were stained with CD80 antibodies and with a suitable counterstaining. As a control, CD80 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 CD80. Peritoneal macrophages from C57BL/6 mice were stained with CD80 antibodies and with a suitable counterstaining. As a control, CD80 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 CD80 (2D10). 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 CD80 (2D10). 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 CD80 Antibody, anti-human

Overview

CD80, also known as B7-1, is a 262 amino acid long 60 kDa molecule and a member of the immunoglobulin superfamily. Together with CD86 (B7-2) it belongs to the B7 family of costimulatory molecules. CD80 is expressed on activated B cells, dendritic cells, and monocytes/ macrophages. The interaction of CD80 with its ligands CD28 and CD152 (CTLA-4) plays a critical role in induction and regulation of immune responses. Binding of CD80 to CD28, which is constitutively expressed on T cells, provides a costimulatory signal and induces T cell proliferation and cytokine production. In contrast, binding to CTLA-4 strongly inhibits proliferation and interleukin 2 (IL-2) secretion by T cells.

Alternative names

B7, B7-1, BB1, CD28LG, CD28LG1, LAB7

Detailed product information

Technical specifications

Clone	2D10
Clonality	monoclonal
Isotype	mouse IgG1κ
Isotype control	Isotype Control Antibody, mouse IgG1
Host	mouse
Type of antibody	Primary antibodies
Species	human, non-human primate
Cross-reactivity	rhesus monkey ( Macaca mulatta )
Antigen	CD80
Alternative names of antigen	B7, B7-1, BB1, CD28LG, CD28LG1, LAB7
Molecular mass of antigen [kDa]	29
Distribution of antigen	B cells, dendritic cells, macrophages, monocytes, Langerhans cells, T cells
Entrez Gene ID	941
RRID	AB_2733839, AB_2801992, AB_2801971, AB_2857834, AB_2857831, AB_2659263, AB_2733838

Resources for CD80 Antibody, anti-human

Certificates

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References for CD80 Antibody, anti-human

Publications

Krummel, M. F. et al. (1995) CD28 and CTLA-4 have opposing effects on the response of T cells to stimulation. J. Exp. Med. 182(2): 459-465
Linsley, P. S. et al. (1991) Binding of the B cell activation antigen B7 to CD28 costimulates T cell proliferation and interleukin 2 mRNA accumulation. J. Exp. Med. 173(3): 721-730

Applications

Extended validation