Clone:
REA306
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC, MC
Alternative names:
CEACAM8, CD67, CGM6, NCA-95

Extended validation for CD66b 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 REA306
G10F5++
80H3-
Cells were incubated with an excess of purified unconjugated CD66b (REA306) 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 CD66b. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD66b antibodies and plotted against the side scatter. As a control, CD66b 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 CD66b. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD66b antibodies and plotted against the side scatter. As a control, CD66b 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 CD66b. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD66b antibodies and plotted against the side scatter. As a control, CD66b 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 CD66b. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD66b antibodies and plotted against the side scatter. As a control, CD66b 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 CD66b. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD66b antibodies and plotted against the side scatter. As a control, CD66b 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 CD66b. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD66b antibodies and plotted against the side scatter. As a control, CD66b 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 CD66b (REA306). 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 CD66b (REA306). 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 CD66b (REA306). 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 CD66b Antibody, anti-human, REAfinity™

Overview

Clone REA306 recognizes the human CD66b antigen, a glycosylphosphatidylinositol (GPI)–linked protein, which is also known as carcinoembryonic antigen-related cell adhesion molecule 8 (CEACAM8). CD66b is exclusively expressed on neutrophils and eosinophils and on leukocytes of chronic myeloid leukemia patients and bone marrow. Under normal conditions, neutrophils have minimal expression of CD66b, but incubation
in vitro
with inflammatory agonists rapidly increases this expression. Furthermore, expression of CD66b in eosinophils is enhanced
in vivo
in patients with a helminth infection, specifically with
Schistosoma mansoni
. The eosinophil expression of CD66b is also enhanced
in vitro
by incubation with activated lymphocytes or by brief incubation with interleukin 5 (IL-5). Crosslinking of CD66b on peripheral blood neutrophils mediates the release of IL-8 from intracellular storage.
Additional information: Clone REA306 displays negligible binding to Fc receptors.

Alternative names

CEACAM8, CD67, CGM6, NCA-95

Detailed product information

Technical specifications

CloneREA306
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD66b
Alternative names of antigenCEACAM8, CD67, CGM6, NCA-95
Molecular mass of antigen [kDa]32
Distribution of antigengranulocytes, eosinophils, neutrophils, bone marrow
Entrez Gene ID1088
RRIDAB_2733400, AB_2733532, AB_2733533, AB_2751860, AB_2751829, AB_2752011, AB_2751981, AB_2811418, AB_2811406, AB_2802064, AB_2802056, AB_2658994, AB_2733399

Resources for CD66b Antibody, anti-human, REAfinity™

References for CD66b Antibody, anti-human, REAfinity™

Publications

  1. Yoon, J. et al. (2007) CD66b regulates adhesion and activation of human eosinophils. J. Immunol. 179(12): 8454-8462
  2. Molgora, M. et al. (2017) IL-1R8 is a checkpoint in NK cells regulating anti-tumour and anti-viral activity. Nature 551(7678): 110-114
  3. Eades-Perner, A. M. et al. (1998) Mice transgenic for the human CGM6 gene express its product, the granulocyte marker CD66b, exclusively in granulocytes. Blood 91(2): 663-672
  4. Berling, B. et al. (1990) Cloning of a carcinoembryonic antigen gene family member expressed in leukocytes of chronic myeloid leukemia patients and bone marrow. Cancer Res. 50(20): 6534-6539
  5. Pearl, M. L. et al. (2014) Prognostic analysis of invasive circulating tumor cells (iCTCs) in epithelial ovarian cancer. Gynecol. Oncol. 134(3): 581-590
  6. Liakopoulos, V. et al. (2018) Hemodialysis-related changes in phenotypical features of monocytes. Sci Rep 8(1): 13964

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