Clone:
REA219
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC
Alternative names:
LILRA2, ILT1, LIR-7

Extended validation for CD85h (ILT1) 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 REA219
24++
135.4++
Cells were incubated with an excess of purified unconjugated CD85h (ILT1) (REA219) 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 CD85h (ILT1). Splenocytes from C57BL/6 mice were stained with a suitable counterstaining and then fixed and permeabilized using the FoxP3 Staining Buffer Set. Cells were then stained with CD85h (ILT1) antibodies. As a control, CD85h (ILT1) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals. 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 CD85h (ILT1). Splenocytes from C57BL/6 mice were stained with a suitable counterstaining and then fixed and permeabilized using the FoxP3 Staining Buffer Set. Cells were then stained with CD85h (ILT1) antibodies. As a control, CD85h (ILT1) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals. 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 CD85h (ILT1). Splenocytes from C57BL/6 mice were stained with a suitable counterstaining and then fixed and permeabilized using the FoxP3 Staining Buffer Set. Cells were then stained with CD85h (ILT1) antibodies. As a control, CD85h (ILT1) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for CD85h (ILT1). Splenocytes from C57BL/6 mice were stained with a suitable counterstaining and then fixed and permeabilized using the FoxP3 Staining Buffer Set. Cells were then stained with CD85h (ILT1) antibodies. As a control, CD85h (ILT1) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals. 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 CD85h (ILT1) (REA219). 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 CD85h (ILT1) (REA219). 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 CD85h (ILT1) (REA219). 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 CD85h (ILT1) Antibody, anti-human, REAfinity™

Overview

Clone REA219 recognizes CD85h (ILT1), a member of the immunoglobulin-like transcripts (ILT) family of proteins. ILT family members compose of both activating and inhibitory receptors, where CD85h serves as an activating receptor. In contrast to inhibitory members, CD85h posesses a positively charged amino acid in its trasmembrane domain and a short cytoplasmic domain which is devoid of sequence motifs involved in signal transduction. To transduce activating signal, ILT1 forms a multiprotein complex with FCεRIγ. Expression of ILT1 is found on the surface of monocytes/macrophages, dendritic cells, and on a small subset of NK cells.
Additional information: Clone REA219 displays negligible binding to Fc receptors.

Alternative names

LILRA2, ILT1, LIR-7

Detailed product information

Technical specifications

CloneREA219
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD85h (ILT1)
Alternative names of antigenLILRA2, ILT1, LIR-7
Distribution of antigendendritic cells, macrophages, monocytes
RRIDAB_2659363, AB_2659364, AB_2659365, AB_2659366, AB_2659367, AB_2659368, AB_2659369, AB_2659370, AB_2659371, AB_2659372, AB_2659373, AB_2659362

References for CD85h (ILT1) Antibody, anti-human, REAfinity™

Publications

  1. Nakajima, H. et al. (1999) Cutting edge: human myeloid cells express an activating ILT receptor (ILT1) that associates with Fc receptor γ-chain. J. Immunol. 162: 5-8
  2. Chen, Y. et al. (2009) Crystal structure of myeloid cell activating receptor leukocyte Ig-like receptor A2 (LILRA2/ILT1/LIR-7) domain swapped dimer: molecular basis for its non-binding to MHC complexes. J. Mol. Biol. 386(3): 841-853

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