Clone:
REA442
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
ALCAM, MEMD

Extended validation for CD166 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 REA442
3A6++
Cells were incubated with an excess of purified unconjugated CD166 (REA442) 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 CD166. Human peripheral blood mononuclear cells (PBMCs) were stained with CD166antibodies and with a suitable counterstaining. As a control, CD166antibody 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 CD166. Human peripheral blood mononuclear cells (PBMCs) were stained with CD166antibodies and with a suitable counterstaining. As a control, CD166antibody 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 CD166. Human peripheral blood mononuclear cells (PBMCs) were stained with CD166antibodies and with a suitable counterstaining. As a control, CD166antibody 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 CD166. Human peripheral blood mononuclear cells (PBMCs) were stained with CD166antibodies and with a suitable counterstaining. As a control, CD166antibody 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 CD166. Human peripheral blood mononuclear cells (PBMCs) were stained with CD166antibodies and with a suitable counterstaining. As a control, CD166antibody 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 CD166. Human peripheral blood mononuclear cells (PBMCs) were stained with CD166antibodies and with a suitable counterstaining. As a control, CD166antibody 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 CD166. Human peripheral blood mononuclear cells (PBMCs) were stained with CD166antibodies and with a suitable counterstaining. As a control, CD166antibody 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 CD166. Human peripheral blood mononuclear cells (PBMCs) were stained with CD166antibodies and with a suitable counterstaining. As a control, CD166antibody 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 CD166 (REA442). 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 CD166 (REA442). 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 CD166 (REA442). 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 CD166 (REA442). 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 CD166 (REA442). 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 CD166 Antibody, anti-human, REAfinity™

Overview

Clone REA442 recognizes the human CD166 antigen, a single-pass type I membrane protein also known as activated leukocyte cell adhesion molecule (ALCAM). CD166, a member of the immunoglobulin superfamily and a ligand for the lymphocyte antigen CD6, mediates homophilic and heterophilic adhesion. It is expressed on activated leukocytes T cells, B cells, monocytes, hematopoietic stem cells (HSCs), metastasizing melanoma, neuronal cells, endothelial cells, hematopoiesis-supporting osteoblastic cell lines, and mesenchymal stem cells (MSCs). CD166 expression is pathologically correlated with aggressive disease in a variety of cancers including melanoma, prostate, breast, ovarian, esophageal, and bladder cancers. It is also a universal functional marker of murine and human HSCs and osteoblasts within the hematopoietic niche and it is involved in modulating HSC-niche interactions and HSC fate.
Additional information: Clone REA442 displays negligible binding to Fc receptors.

Alternative names

ALCAM, MEMD

Detailed product information

Technical specifications

CloneREA442
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD166
Alternative names of antigenALCAM, MEMD
Molecular mass of antigen [kDa]62
Distribution of antigenosteoblasts, T cells, B cells, monocytes, endothelial cells, stem cells, hematopoietic stem and progenitor cells, melanoma, mesenchymal stem cells, melanoma
Entrez Gene ID214
RRIDAB_2733774, AB_2751861, AB_2751830, AB_2819751, AB_2819746, AB_2655530, AB_2655531, AB_2733773

Resources for CD166 Antibody, anti-human, REAfinity™

Certificates

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References for CD166 Antibody, anti-human, REAfinity™

Publications

  1. Chitteti, B. R. et al. (2014) CD166 regulates human and murine hematopoietic stem cells and the hematopoietic niche. Blood 124(4): 519-529
  2. Degen, W. G. et al. (1998) MEMD, a new cell adhesion molecule in metastasizing human melanoma cell lines, is identical to ALCAM (activated leukocyte cell adhesion molecule). Am. J. Pathol. 152(3): 805-813
  3. Ni, C. et al. (2013) Prognostic value of CD166 expression in cancers of the digestive system: a systematic review and meta-analysis. PLoS One 8(8): e70958

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