CD85a (ILT5) Antibody, anti-human, REAfinity™

Name: CD85a (ILT5) Antibody, anti-human, REAfinity™
Availability: InStock

CD85a (ILT5) Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA207

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC

Alternative names:

LILRB3, HL9, ILT5, LILRA6, LIR-3, PIRB

Flow cytometry applications forCD85a (ILT5) Antibody, anti-human, REAfinity™

APC
APC-Vio 770
Biotin
FITC
PE
PE-Vio 770
PerCP-Vio 700
Vio Bright R720
VioBlue

Conjugate:

APC

Recommended dilution:

1:11

Remark:

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

Tested:

QC tested

Products used:

130-101-665, 130-101-733

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:11)

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD85a (ILT5) antibodies as well as with CD14 antibodies 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 CD85a (ILT5) Antibody, anti-human, 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 REA207
MKT5.1	++
222821	+
Cells were incubated with an excess of purified unconjugated CD85a (ILT5) (REA207) 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 CD85a (ILT5). Human peripheral blood mononuclear cells (PBMCs) were stained with CD85a (ILT5) antibodies and with a suitable counterstaining. As a control, CD85a (ILT5) 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 CD85a (ILT5). Human peripheral blood mononuclear cells (PBMCs) were stained with CD85a (ILT5) antibodies and with a suitable counterstaining. As a control, CD85a (ILT5) 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 CD85a (ILT5). Human peripheral blood mononuclear cells (PBMCs) were stained with CD85a (ILT5) antibodies and with a suitable counterstaining. As a control, CD85a (ILT5) 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 CD85a (ILT5) (REA207). 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 CD85a (ILT5) (REA207). 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 CD85a (ILT5) Antibody, anti-human, REAfinity™

Overview

Clone REA207 recognizes the CD85a antigen, a immunoglobulin superfamily transmembrane glycoprotein also known as leukocyte immunoglobulin-like receptor member 3 (LIR-3) or immunoglobulin-like transcript 5 (ILT5). CD85a is preferentially expressed by monocytes, macrophages, dendritic cells, and granulocytes. It provides inhibitory signals through interaction with MHC class I molecules and becomes activated upon co-ligation of CD85a and immune receptors, such as CD32 and the B cell receptor. CD85a down-regulates antigen-induced B cell–activation by recruiting phosphatases to its immunoreceptor tyrosine-based inhibitory motifs.

Additional information: Clone REA207 displays negligible binding to Fc receptors.

Alternative names

LILRB3, HL9, ILT5, LILRA6, LIR-3, PIRB

Detailed product information

Technical specifications

Clone	REA207
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody (S), human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	human
Antigen	CD85a (ILT5)
Alternative names of antigen	LILRB3, HL9, ILT5, LILRA6, LIR-3, PIRB
Distribution of antigen	dendritic cells, granulocytes, macrophages
Entrez Gene ID	11025
RRID	AB_2659333, AB_2659334, AB_2659335, AB_2659336, AB_2659337, AB_2659338, AB_2659339, AB_2659340, AB_2659341, AB_2659342, AB_2659343, AB_2659344, AB_2659345, AB_2922252, AB_2922219, AB_2928572, AB_2928571, AB_2659332

Resources for CD85a (ILT5) Antibody, anti-human, REAfinity™

Certificates

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References for CD85a (ILT5) Antibody, anti-human, REAfinity™

Publications

Borges, L. et al. (1997) A family of human lymphoid and myeloid Ig-like receptors, some of which bind to MHC class I molecules. J Immunol 159: 5192-5196
Ma, G. et al. (2011) Paired immunoglobin-like receptor-B regulates the suppressive function and fate of myeloid-derived suppressor cells. Immunity 34(3): 385-395
Colonna, M. et al. (1999) Inhibitory and activating receptors involved in immune surveillance by human NK and myeloid cells. J. Leukoc. Biol. 66: 718-722

Applications

Extended validation