Clone:
REA184
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
MC, FC
Alternative names:
LILRB2, ILT-4, LIR2, MIR-10

Extended validation for CD85d (ILT4) 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 REA184
42D1++
287219-
Cells were incubated with an excess of purified unconjugated CD85d (ILT4) (REA184) 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 CD85d (ILT4). Human peripheral blood mononuclear cells (PBMCs) were stained with CD85d (ILT4) antibodies and with a suitable counterstaining. As a control, CD85d (ILT4) 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 CD85d (ILT4). Human peripheral blood mononuclear cells (PBMCs) were stained with CD85d (ILT4) antibodies and with a suitable counterstaining. As a control, CD85d (ILT4) 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 CD85d (ILT4). Human peripheral blood mononuclear cells (PBMCs) were stained with CD85d (ILT4) antibodies and with a suitable counterstaining. As a control, CD85d (ILT4) 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 CD85d (ILT4). Human peripheral blood mononuclear cells (PBMCs) were stained with CD85d (ILT4) antibodies and with a suitable counterstaining. As a control, CD85d (ILT4) 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 CD85d (ILT4) (REA184). 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 CD85d (ILT4) (REA184). 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 CD85d (ILT4) (REA184). 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 CD85d (ILT4) Antibody, anti-human, REAfinity™

Overview

Clone REA184 recognizes CD85d, also known as immunoglobulin-like transcript 4 (ILT4) or leukocyte Ig-like receptor 2 (LIR2). CD85d is an inhibitory immunoreceptor, expressed mainly on myelomonocytic lineage cells.
CD85d has four tandem Ig-like extracellular domains and three immunoreceptor tyrosine-based inhibitory receptor motifs in its cytoplasmic tails, which recruit the tyrosine phosphatase Src homology 2 domain containing phosphatase 1 (SHP-1). It has been shown that CD85d interacts with HLA-A, -B, -G1, and -E molecules as ligands. HLA-G expression is restricted to trophoblasts and thus interaction of CD85d with HLA-G is suggested to inhibit placental leukocytes, contributing to maternal tolerance of the fetal semiallograft.
Additional information: Clone REA184 displays negligible binding to Fc receptors.

Alternative names

LILRB2, ILT-4, LIR2, MIR-10

Detailed product information

Technical specifications

CloneREA184
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD85d (ILT4)
Alternative names of antigenLILRB2, ILT-4, LIR2, MIR-10
Molecular mass of antigen [kDa]63
Distribution of antigengranulocytes, monocytes
Entrez Gene ID10288
RRIDAB_2659347, AB_2659346, AB_2659348, AB_2659349

References for CD85d (ILT4) Antibody, anti-human, REAfinity™

Publications

  1. Shiroishi, M. et al. (2003) Human inhibitory receptors Ig-like transcript 2 (ILT2) and ILT4 compete with CD8 for MHC class I binding and bind preferentially to HLA-G. Proc. Natl. Acad. Sci. U.S.A. 100(15): 8856-8861
  2. Shiroishi, M. et al. (2006) Structural basis for recognition of the nonclassical MHC molecule HLA-G by the leukocyte Ig-like receptor B2 (LILRB2/LIR2/ILT4/CD85d). Proc. Natl. Acad. Sci. U.S.A. 103: 16412-16417
  3. Ristich, V. et al. (2007) Mechanisms of prolongation of allograft survival by HLA-G/ILT4-modified dendritic cells. Hum. Immunol. 68(4): 264-271

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