Clone:
REA428
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
BGP-1, CEACAM5, CEACAM3, CEACAM6, CGM1, NCA, CEA, CEACAM1

Extended validation for CD66acde 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 REA428
B1.1/CD66++
REA414+
Cells were incubated with an excess of purified unconjugated CD66acde (REA428) 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 CD66acde. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD66acde antibodies and with a suitable counterstaining. As a control, CD66acde 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 CD66acde. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD66acde antibodies and with a suitable counterstaining. As a control, CD66acde 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 CD66acde. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD66acde antibodies and with a suitable counterstaining. As a control, CD66acde 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 CD66acde. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD66acde antibodies and with a suitable counterstaining. As a control, CD66acde 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 CD66acde (REA428). 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 CD66acde (REA428). 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 CD66acde (REA428). 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 CD66acde Antibody, anti-human, REAfinity™

Overview

Clone REA428 recognizes an epitope shared by the human CD66a, c, d, and e antigens, which are also known as the human carcinoembryonic antigen (CEA) family. The CEA family has 7 genes belonging to the CEACAM subgroup. These subgroup members are mainly associated with the cell membrane and show a complex expression pattern in normal and cancerous tissues. CD66a (CEACAM1) is an adhesion molecule that is involved in many immune responses associated with infection, inflammation, and cancer. It interacts homophilically with CD66a and heterophilically with CD66e (CEACAM5), but not with other CEACAM proteins. CD66a is expressed on a variety of cells, e.g., some epithelial cells, melanoma, and activated lymphocytes. Within the hematopoietic system, CD66c (CEACAM6) expression is limited to granulocytes and its precursors, where it serves homotypic and heterotypic adhesion and Ca2
+
mediated signaling. It is markedly upregulated from intracellular stores after activatio and is also found in epithelia of various organs. CD66c overexpression is found in a wide variety of epithelial cancer types such as lung, breast, colorectal, and hepatocellular carcinomas. CD66d (CEACAM3) is a major granulocyte receptor mediating recognition and efficient opsonin-independent phagocytosis of CEACAM-binding microorganisms, thus playing an important role in the clearance of pathogens by the innate immune system. CD66e (CEACAM5) plays a role in cell adhesion and in intracellular signaling. It is known to be overexpressed in a majority of carcinomas, including those of the gastrointestinal tract, the respiratory and genitourinary systems, and breast cancer.
Additional information: Clone REA428 displays negligible binding to Fc receptors.

Alternative names

BGP-1, CEACAM5, CEACAM3, CEACAM6, CGM1, NCA, CEA, CEACAM1

Detailed product information

Technical specifications

CloneREA428
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD66acde
Alternative names of antigenBGP-1, CEACAM5, CEACAM3, CEACAM6, CGM1, NCA, CEA, CEACAM1
Molecular mass of antigen [kDa]45(average of subunits)
Distribution of antigenepithelial cells, granulocytes, lymphocytes, melanoma
Entrez Gene ID634, 4680, 1084, 1048
RRIDAB_2857753, AB_2658970, AB_2658971, AB_2658972, AB_2658973, AB_2658976, AB_2658977, AB_2658978, AB_2658979, AB_2658980, AB_2658981, AB_2857765

References for CD66acde Antibody, anti-human, REAfinity™

Publications

  1. Markel, G. et al. (2002) CD66a interactions between human melanoma and NK cells: a novel class I MHC-independent inhibitory mechanism of cytotoxicity. J. Immunol. 168(6): 2803-2810
  2. Streichert, T. et al. (2001) The microbial receptor CEACAM3 is linked to the calprotectin complex in granulocytes. Biochem. Biophys. Res. Commun. 289(1): 191-197
  3. Guillaume, N. et al. (2011) CD66c expression in B-cell acute lymphoblastic leukemia: strength and weakness. Int J Lab Hematol 33(1): 92-96

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