Clone:
REA753
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC, MC
Alternative names:
PROM1, AC133, CORD12, MCDR2, MSTP061, PROML1, RP41, STGD4

Extended validation for CD133/1 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 REA753
293C3-
AC133++
AC141-
clone 7++
EMK08++
REA816-
REA820-
REAL233++
S16015F-
S16016B-
S16016E-
TMP4-
W6B3C1++
Cells were incubated with an excess of purified unconjugated CD133/1 (REA753) 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 CD133/1. Human peripheral blood mononuclear cells (PBMCs) were stained with CD133/1 antibodies and with a suitable counterstaining. As a control, CD133/1 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD45+ cells were pregated for the analysis. 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 CD133/1. Human peripheral blood mononuclear cells (PBMCs) were stained with CD133/1 antibodies and with a suitable counterstaining. As a control, CD133/1 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD45+ cells were pregated for the analysis. 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 CD133/1. Human peripheral blood mononuclear cells (PBMCs) were stained with CD133/1 antibodies and with a suitable counterstaining. As a control, CD133/1 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD45+ cells were pregated for the analysis. 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 CD133/1. Human peripheral blood mononuclear cells (PBMCs) were stained with CD133/1 antibodies and with a suitable counterstaining. As a control, CD133/1 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD45+ cells were pregated for the analysis. 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 CD133/1. Human peripheral blood mononuclear cells (PBMCs) were stained with CD133/1 antibodies and with a suitable counterstaining. As a control, CD133/1 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD45+ cells were pregated for the analysis. 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 CD133/1. Human peripheral blood mononuclear cells (PBMCs) were stained with CD133/1 antibodies and with a suitable counterstaining. As a control, CD133/1 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD45+ cells were pregated for the analysis. 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 CD133/1. Human peripheral blood mononuclear cells (PBMCs) were stained with CD133/1 antibodies and with a suitable counterstaining. As a control, CD133/1 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD45+ cells were pregated for the analysis. 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 CD133/1. Human peripheral blood mononuclear cells (PBMCs) were stained with CD133/1 antibodies and with a suitable counterstaining. As a control, CD133/1 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD45+ cells were pregated for the analysis. 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 CD133/1. Human peripheral blood mononuclear cells (PBMCs) were stained with CD133/1 antibodies and with a suitable counterstaining. As a control, CD133/1 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD45+ cells were pregated for the analysis. 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 CD133/1 (REA753). 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 CD133/1 (REA753). 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 CD133/1 (REA753). 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 CD133/1 Antibody, anti-human, REAfinity™

Overview

Clone REA753 recognizes the epitope 1 of the human CD133 antigen (CD133/1). This molecule is frequently found on multipotent progenitor cells, including immature hematopoietic stem and progenitor cells. In the hematopoietic system, CD133 is expressed on a small portion of CD34
cells as well as on a subset of CD34
bright
stem and progenitor cells in human fetal liver, bone marrow, cord blood, and peripheral blood. CD133 has also been found to be expressed on circulating endothelial progenitor cells, fetal neural stem cells, other tissue-specific stem cells, such as renal, prostate, and corneal stem cells, cancer stem cells from tumor tissues, as well as ES and iPS cell–derived cells.
Additional information: Clone REA753 displays negligible binding to Fc receptors.

Alternative names

PROM1, AC133, CORD12, MCDR2, MSTP061, PROML1, RP41, STGD4

Detailed product information

Technical specifications

CloneREA753
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD133/1
Alternative names of antigenPROM1, AC133, CORD12, MCDR2, MSTP061, PROML1, RP41, STGD4
Molecular mass of antigen [kDa]95
Distribution of antigenendothelial cells, epithelial cells, red blood cells, ES and iPS cells, hematopoietic stem and progenitor cells, brain, heart, kidney, liver, lung, pancreas, placenta
Entrez Gene ID8842
RRIDAB_2654886, AB_2654887, AB_2654888, AB_2654889, AB_2654890, AB_2654891, AB_2654892, AB_2654893, AB_2654894, AB_2654895, AB_2654896, AB_2751051, AB_2751049, AB_2654897, AB_2654898, AB_2811644, AB_2654885

References for CD133/1 Antibody, anti-human, REAfinity™

Publications

  1. Giebel, B. et al. (2004) Segregation of lipid raft markers including CD133 in polarized human hematopoietic stem and progenitor cells. Blood 104(8): 2332-2338
  2. Ingram, D. A. et al. (2004) Identification of a novel hierarchy of endothelial progenitor cells using human peripheral and umbilical cord blood. Blood 104(9): 2752-2760
  3. Miraglia, S. et al. (1997) A novel five-transmembrane hematopoietic stem cell antigen: isolation, characterization, and molecular cloning. Blood 90(12): 5013-5021
  4. Wagner, W. et al. (2004) Molecular evidence for stem cell function of the slow-dividing fraction among human hematopoietic progenitor cells by genome-wide analysis. Blood 104(3): 675-686
  5. Yin, A. H. et al. (1997) AC133, a novel marker for human hematopoietic stem and progenitor cells. Blood 90: 5002-5012
  6. Gallacher, L. et al. (2000)
    Isolation and characterization of human CD34
    Lin
    and CD34
    +
    Lin
    hematopoietic stem cells using cell surface markers AC133 and CD7.
    Blood 95(ARVO Annual Meeting Abstract): 2813-2820
  7. Gehling, U. M. et al. (2000)
    In vitro
    differentiation of endothelial cells from AC133-positive progenitor cells.
    Blood 95: 3106-3112
  8. Peichev, M. et al. (2000)
    Expression of VEGFR-2 and AC133 by circulating human CD34
    +
    cells identifies a population of functional endothelial precursors.
    Blood 95: 952-958
  9. Cummings, B. J. et al. (2005) Human neural stem cells differentiate and promote locometer recovery in spinal cord-injured mice. Proc. Natl. Acad. Sci. U.S.A. 102: 14069-14074
  10. Galic, Z. et al. (2006) T lineage differentiation from human embryonic stem cells. Proc. Natl. Acad. Sci. U.S.A. 103: 11742-11747
  11. Uchida, N. et al. (2000) Direct isolation of human central nervous system stem cells. Proc. Natl. Acad. Sci. U.S.A. 97: 14720-14725
  12. Weigmann, A. et al. (1997) Prominin, a novel microvilli-specific polytopic membrane proteine of the apical surface of epithelial cells, is targeted to plasmalemmal protrusions of non-epithelial cells. Proc. Natl. Acad. Sci. U.S.A. 94: 12425-12430
  13. Estes, M. L. et al. (2010) Application of polychromatic flow cytometry to identify novel subsets of circulating cells with angiogenic potential. Cytometry A 77(9): 831-839
  14. Park, T. S. et al. (2013) Efficient and simultaneous generation of hematopoietic and vascular progenitors from human induced pluripotent stem cells. Cytometry A 83(1): 114-126
  15. Bussolati, B. et al. (2005) Isolation of renal progenitor cells from adult human kidney. Am. J. Pathol. 166: 545-555
  16. Bühring, H. J. et al. (1999) Expression of novel surface antigens on early hematopoietic cells. Ann. N. Y. Acad. Sci. 872: 25-39
  17. Immervoll, H. et al. (2008) Expression of the "stem cell marker" CD133 in pancreas and pancreatic ductal adenocarcinomas. BMC Cancer 8: 48
  18. Wong, C. K. E. et al. (2012) Levels of a subpopulation of platelets, but not circulating endothelial cells, predict early treatment failure in prostate cancer patients after prostatectomy. Br. J. Cancer 107(9): 1564-1573
  19. Richardson, G. et al. (2004) CD133, a novel marker for human prostatic epithelial stem cells. J. Cell. Sci. 117: 3539-3545
  20. Walton, R. M. et al. (2013)
    Postnatal neural precursor cell regions in the rostral subventricular zone, hippocampal subgranular zone and cerebellum of the dog (
    Canis lupus familiaris
    ).
    Histochem. Cell Biol. 139(3): 415-429
  21. Fusi, A. et al. (2011) Expression of the stem cell markers nestin and CD133 on circulating melanoma cells. J. Invest. Dermatol. 131(2): 487-494
  22. Thill, M. et al. (2004)
    Identification of a population of CD133
    +
    precursor cells in the stroma of human cornea.
    Invest. Ophthalmol. Vis. Sci. 45: 3519
  23. Fonseca, A. V. et al. (2010) Polarization and migration of hematopoietic stem and progenitor cells rely on the J. Biol. Chem. 285(41): 31661-31671
  24. Mak, A. B. et al. (2011) CD133 protein N-glycosylation processing contributes to cell surface recognition of the primitive cell marker AC133 epitope. J. Biol. Chem. 286(47): 41046-41056
  25. Maxwell, S. A. et al. (2009) 14-3-3zeta J. Biol. Chem. 284(33): 22379-22389
  26. Piechaczek, C. (2001) CD133. J. Biol. Regul. Homeost. Agents 15: 101-102
  27. Margolis, D. J. et al. (2009) Phase I study of H5.020CMV.PDGF-beta to treat venous leg ulcer disease. Mol. Ther. 17(10): 1822-1829
  28. Duda, D. G. et al. (2007) A protocol for phenotypic detection and enumeration of circulating endothelial cells and circulating progenitor cells in human blood. Nat. Protoc. 2(4): 805-810
  29. Clark, P. A. et al. (2012) Activation of multiple ERBB family receptors mediates glioblastoma cancer stem-like cell resistance to EGFR-targeted inhibition. Neoplasia 14(5): 420-428
  30. Lin, L. et al. (2009) The STAT3 inhibitor NSC 74859 is effective in hepatocellular cancers with Oncogene 28(7): 961-972
  31. Ferro, F. et al. (2012) Dental pulp stem cells differentiation reveals new insights in Oct4A dynamics. PLoS One 7(7): e41774
  32. Fredebohm, J. et al. (2012) Establishment and characterization of a highly tumourigenic and cancer stem cell enriched pancreatic cancer cell line as a well defined model system. PLoS One 7(11): e48503
  33. Balasubramanian, P. et al. (2013)
    AQP9 expression in glioblastoma multiforme tumors is limited to a small population of astrocytic cells and CD15
    +
    /CalB
    +
    leukocytes.
    PLoS One 8(9): e75764
  34. Metsuyanim, S. et al. (2009) Expression of stem cell markers in the human fetal kidney. PLoS One 4(68): e6709
  35. Qi, Y. et al. (2012) Inhaled NO contributes to lung repair in piglets with acute respiratory distress syndrome via increasing circulating endothelial progenitor cells. PLoS One 7(3): e33859
  36. Ye, X. et al. (2013) The effect of Heparin-VEGF multilayer on the biocompatibility of decellularized aortic valve with platelet and endothelial progenitor cells. PLoS One 8(1): e54622
  37. Hu, Y. et al. (2013) Tumor-specific chromosome mis-segregation controls cancer plasticity by maintaining tumor heterogeneity. PLoS One 8(11): e80898
  38. de Wynter, E. A. et al. (1998)
    CD34
    +
    AC133
    +
    cells isolated from cord blood are highly enriched in long-term culture-initiating cells, NOD/SCID-repopulating cells and dendritic cell progenitors.
    Stem Cells 16: 387-396
  39. Alvarez, D. F. et al. (2008) Lung microvascular endothelium is enriched with progenitor cells that exhibit vasculogenic capacity. Am. J. Physiol. Lung Cell Mol. Physiol. 294(3): L419-L430
  40. Rheinbay, E. et al. (2013) An aberrant transcription factor network essential for Wnt signaling and stem cell maintenance in glioblastoma. Cell Rep 3(5): 1567-1579
  41. Behbod, F. et al. (2009)
    An intraductal human-in-mouse transplantation model mimics the subtypes of ductal carcinoma
    in situ
    .
    Breast Cancer Res. 11(5): R66
  42. Steinmetz, N. F. et al. (2011) Two domains of vimentin are expressed on the surface of lymph node, bone and brain metastatic prostate cancer lines along with the putative stem cell marker proteins CD44 and CD133. Cancers (Basel) 3(3): 2870-2885
  43. Liu, H. et al. (2013) Single-cell clones of liver cancer stem cells have the potential of differentiating into different types of tumor cells. Cell Death Dis 4: e857
  44. Czyz, M. et al. (2013) Parthenolide reduces the frequency of ABCB5-positive cells and clonogenic capacity of melanoma cells from anchorage independent melanospheres. Cancer Biol. Ther. 14(2): 135-145
  45. Tsang, H. et al. (2013) Role of asymmetric methylarginine and connexin 43 in the regulation of pulmonary endothelial function. Pulm Circ 3(3): 675-691
  46. Sun, Y. et al. (2014) Slug overexpression induces stemness and promotes hepatocellular carcinoma cell invasion and metastasis. Oncol Lett 7(6): 1936-1940
  47. Schiffer, D. et al. (2014) Stem cell niches in glioblastoma: a neuropathological view. Biomed Res Int 2014: 725921

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