Clone:
REA113
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC, MC
Alternative names:
KLRD1, Kp43

Extended validation for CD94 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 REA113
HP-3D9++
DX22++
Cells were incubated with an excess of purified unconjugated CD94 (REA113) 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 CD94. Human peripheral blood mononuclear cells (PBMCs) were stained with CD94 antibodies and with a suitable counterstaining. As a control, CD94 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 CD94. Human peripheral blood mononuclear cells (PBMCs) were stained with CD94 antibodies and with a suitable counterstaining. As a control, CD94 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 CD94. Human peripheral blood mononuclear cells (PBMCs) were stained with CD94 antibodies and with a suitable counterstaining. As a control, CD94 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 CD94. Human peripheral blood mononuclear cells (PBMCs) were stained with CD94 antibodies and with a suitable counterstaining. As a control, CD94 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 CD94. Human peripheral blood mononuclear cells (PBMCs) were stained with CD94 antibodies and with a suitable counterstaining. As a control, CD94 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 CD94 (REA113). 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 CD94 (REA113). 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 CD94 (REA113). 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 CD94 Antibody, anti-human, REAfinity™

Overview

The clone REA113 recognizes CD94, an approximately 45 kDa, single-pass type II transmembrane protein, which belongs to the C-type lectin superfamily. CD94 associates covalently with NKG2 family members and forms disulfide bonded heterodimers. CD94-NKG2 receptors bind HLA-E, the non-classical MHC class Ib molecule. Depending on the associated NKG2 member, CD94-NKG2 heterodimer transmits either activating or inhibitory signal. For, e.g, via immunoreceptor tyrosine based-inhibitory motifs (ITIMs) containing NKG2A, CD94-NKG2A heterodimers inhibit NK cell cytotoxicity. Besides NK cells, expression of CD94-NKG2 heterodimers is also shown on NK-T cells and a small percentage of CD8 cells in mice, where the crosslinking of CD94-NKG2 was shown to reduce the level of spontaneous apoptosis.
Additional information: Clone REA113 displays negligible binding to Fc receptors.

Alternative names

KLRD1, Kp43

Detailed product information

Technical specifications

CloneREA113
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
Cross-reactivity
african green monkey (
Chlorocebus aethiops
)
, baboon,
cynomolgus monkey (
Macaca fascicularis
)
, canine
AntigenCD94
Alternative names of antigenKLRD1, Kp43
Molecular mass of antigen [kDa]21
Distribution of antigenNK cells
Entrez Gene ID3824
RRIDAB_2751824, AB_2811494, AB_2811483, AB_2857646, AB_2857615, AB_2659624, AB_2659625, AB_2659626, AB_2659627, AB_2659628, AB_2659629, AB_2659634, AB_2659635, AB_2751855

References for CD94 Antibody, anti-human, REAfinity™

Publications

  1. Gunturi, A. et al. (2003) Preferential survival of CD8 T and NK cells expressing high levels of CD94. J. Immunol. 170: 1737-1745
  2. Braud, V. M. et al. (1998) HLA-E binds to natural killer cell receptors CD94/NKG2A, B and C. Nature 391(6669): 795-799
  3. Brooks, A. G. et al. (1997) NKG2A complexed with CD94 defines a novel inhibitory natural killer cell receptor. J. Exp. Med. 185: 795-800
  4. Jeffery, H. et al. Human intrahepatic ILC2 are IL-13positive amphiregulinpositive and their frequency correlates with model of end stage liver disease score. PLoS One 12(12): e0188649
  5. Wolpert, F. et al. (2015) Interferon-β modulates the innate immune response against glioblastoma initiating cells. PLoS One 10(10): e0139603
  6. Canté-Barrett, K. et al. (2017)
    Loss of CD44
    dim
    expression from early progenitor cells marks T-cell lineage commitment in the human thymus.
    Front Immunol 8: 32

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