Clone:
REA323
Type of antibody:
Recombinant antibodies, Primary antibodies
Isotype:
recombinant human IgG1
Applications:
FC
Alternative names:
HBA71, MIC2, MIC2X, MIC2Y, MSK5X

Extended validation for CD99R 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 REA323
HIT4 (IgM)++
MEM-131+
Cells were incubated with an excess of purified unconjugated CD99R (REA323) 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 CD99R. Human peripheral blood mononuclear cells (PBMCs) were stained with CD99R antibodies and with a suitable counterstaining. As a control, CD99R 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 CD99R. Human peripheral blood mononuclear cells (PBMCs) were stained with CD99R antibodies and with a suitable counterstaining. As a control, CD99R 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 CD99R. Human peripheral blood mononuclear cells (PBMCs) were stained with CD99R antibodies and with a suitable counterstaining. As a control, CD99R 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 CD99R. Human peripheral blood mononuclear cells (PBMCs) were stained with CD99R antibodies and with a suitable counterstaining. As a control, CD99R 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 CD99R (REA323). 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 CD99R (REA323). 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 CD99R (REA323). 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 CD99R Antibody, anti-human, REAfinity™

Overview

Clone REA323 recognizes the human CD99R antigen, an isoform of CD99 with restricted cell distribution. CD99 is a Single-pass type I membrane protein which is also known as E2 molecule or MIC2. It is involved in multiple cellular events including cell adhesion and migration, apoptosis, cell differentiation and regulation of protein trafficking either in physiological or pathological conditions. In contrast to CD99, which is broadly distributed on many cell types, CD99R is expressed only on T cells, NK cells, and monocytes but not on B cells, erythrocytes, or platelets.
Additional information: Clone REA323 displays negligible binding to Fc receptors.

Alternative names

HBA71, MIC2, MIC2X, MIC2Y, MSK5X

Detailed product information

Technical specifications

CloneREA323
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyRecombinant antibodies, Primary antibodies
Specieshuman
AntigenCD99R
Alternative names of antigenHBA71, MIC2, MIC2X, MIC2Y, MSK5X
Molecular mass of antigen [kDa]17
Distribution of antigenmonocytes, NK cells, T cells
Entrez Gene ID4267
RRIDAB_2659705, AB_2659708, AB_2659709, AB_2659704

References for CD99R Antibody, anti-human, REAfinity™

Publications

  1. Dworzak, M. N. et al. (1994) Flow cytometric assessment of human MIC2 expression in bone marrow, thymus, and peripheral blood Blood 83(2): 415-425
  2. Gelin, C. et al. (1989) The E2 antigen, a 32 kd glycoprotein involved in T-cell adhesion processes, is the MIC2 gene product. EMBO J. 8(11): 3253-3259
  3. Wingett, D. et al. (1999) A role for CD99 in T cell activation Cell. Immunol. 193(1): 17-23

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