Clone:
REA141
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC
Alternative names:
LILRB4, ILT3, LIR-5

Extended validation for CD85k (ILT3) 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 clonesOverlap in epitope recognition with REA141
ZM4.1++
ZM3.8++
Cells were incubated with an excess of purified unconjugated CD85k (ILT3) (REA141) 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.
View details
Flow cytometric comparison of different clones for CD85k (ILT3). Human peripheral blood mononuclear cells (PBMCs) were stained with CD85k (ILT3) antibodies and with a suitable counterstaining. As a control, CD85k (ILT3) 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.
View details
Flow cytometric comparison of different clones for CD85k (ILT3). Human peripheral blood mononuclear cells (PBMCs) were stained with CD85k (ILT3) antibodies and with a suitable counterstaining. As a control, CD85k (ILT3) 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.
View details
Flow cytometric comparison of different clones for CD85k (ILT3). Human peripheral blood mononuclear cells (PBMCs) were stained with CD85k (ILT3) antibodies and with a suitable counterstaining. As a control, CD85k (ILT3) 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.
View details
Flow cytometric comparison of different clones for CD85k (ILT3). Human peripheral blood mononuclear cells (PBMCs) were stained with CD85k (ILT3) antibodies and with a suitable counterstaining. As a control, CD85k (ILT3) 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 CD85k (ILT3). Human peripheral blood mononuclear cells (PBMCs) were stained with CD85k (ILT3) antibodies and with a suitable counterstaining. As a control, CD85k (ILT3) 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
View details
Comparison of staining pattern on non-fixed and fixed cells using CD85k (ILT3) (REA141). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using CD85k (ILT3) (REA141). 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 CD85k (ILT3) (REA141). 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 CD85k (ILT3) Antibody, anti-human, REAfinity™

Overview

Clone REA141 recognizes CD85k, a member of regulatory immune receptor family, also known as leukocyte immunoglobulin-like receptor (LILRs/LIRs), immunoglobulin-like transcripts (ILTs), or CD85 family. CD85k is a type I membrane protein with two extracellular Ig-like domains, intracellular immunoreceptor tyrosine-based inhibitory motifs (ITIM) and is expressed on monocytes, dendritic cells, and macrophages. In addition, suppressor T cells induce the expression of CD85k on antigen presenting cells.
Additional information: Clone REA141 displays negligible binding to Fc receptors.

Alternative names

LILRB4, ILT3, LIR-5

Detailed product information

Technical specifications

CloneREA141
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD85k (ILT3)
Alternative names of antigenLILRB4, ILT3, LIR-5
Molecular mass of antigen [kDa]47
Distribution of antigendendritic cells, macrophages, monocytes
Entrez Gene ID11006
RRIDAB_2659388, AB_2659390, AB_2659391, AB_2659392, AB_2659393, AB_2659394, AB_2659395, AB_2659396, AB_2659397, AB_2659389

References for CD85k (ILT3) Antibody, anti-human, REAfinity™

Publications

  1. Colonna, M. et al. (1997) A common inhibitory receptor for major histocompatibility complex class I molecules on human lymphoid and myelomonocytic cells. J. Exp. Med. 186: 1809-1818
  2. Cheng, H. et al. (2011) Crystal structure of leukocyte Ig-like receptor LILRB4 (ILT3/LIR-5/CD85k): A myeloid inhibitory receptor involved in immune tolerance. J. Biol. Chem. 286: 18013-18025
  3. Sommer, A. and Fabri, M. (2015) Vitamin D regulates cytokine patterns secreted by dendritic cells to promote differentiation of IL-22-producing T cells. PLoS One 10(6): e0130395

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