Clone:
293C3
Type of antibody:
Primary antibodies
Isotype:
mouse IgG2bκ
Applications:
FC, MICS, IF, IHC
Alternative names:
PROM1, AC133, CORD12, MCDR2, MSTP061, PROML1, RP41, STGD4

Extended validation for CD133/2 Antibody, anti-human

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 293C3
REA816++
AC133-
AC141++
clone 7-
EMK08-
REA753-
REA820++
REAL233-
S16015F-
S16016B-
S16016E++
TMP4-
W6B3C1-
Cells were incubated with an excess of purified unconjugated CD133/2 (293C3) 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 CD133/2. Human peripheral blood mononuclear cells (PBMCs) were stained with CD133/2 antibodies and with a suitable counterstaining. As a control, CD133/2 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 CD133/2. Human peripheral blood mononuclear cells (PBMCs) were stained with CD133/2 antibodies and with a suitable counterstaining. As a control, CD133/2 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 CD133/2. Human peripheral blood mononuclear cells (PBMCs) were stained with CD133/2 antibodies and with a suitable counterstaining. As a control, CD133/2 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 CD133/2. Human peripheral blood mononuclear cells (PBMCs) were stained with CD133/2 antibodies and with a suitable counterstaining. As a control, CD133/2 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 CD133/2. Human peripheral blood mononuclear cells (PBMCs) were stained with CD133/2 antibodies and with a suitable counterstaining. As a control, CD133/2 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 CD133/2. Human peripheral blood mononuclear cells (PBMCs) were stained with CD133/2 antibodies and with a suitable counterstaining. As a control, CD133/2 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 CD133/2. Human peripheral blood mononuclear cells (PBMCs) were stained with CD133/2 antibodies and with a suitable counterstaining. As a control, CD133/2 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 CD133/2. Human peripheral blood mononuclear cells (PBMCs) were stained with CD133/2 antibodies and with a suitable counterstaining. As a control, CD133/2 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 CD133/2. Human peripheral blood mononuclear cells (PBMCs) were stained with CD133/2 antibodies and with a suitable counterstaining. As a control, CD133/2 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 CD133/2 (293C3). 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 CD133/2 (293C3). 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 CD133/2 (293C3). 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 CD133/2 Antibody, anti-human

Overview

CD133 is a novel 5-transmembrane cell surface antigen with a molecular weight of 117 kDa. CD133/2 (293C3) antibodies recognize epitope 2 of the human CD133 antigen (CD133/2). In the hematopoietic system, CD133 expression is restricted to a subset of CD34
bright
stem and progenitor cells in human fetal liver, bone marrow, cord blood, and peripheral blood. Additionally, CD133 is expressed by a small portion of CD34
cells in these tissues. The CD34
+
CD133
+
cell population, which includes CD34
+
CD38
cells, was shown to be capable of repopulating NOD/SCID mice. Recently, CD133 has also been found to be expressed on circulating endothelial progenitor cells, tissue-specific stem cells, cancer stem cells from tumor tissues, fetal neural stem cells, and ES and iPS cell–derived cells as well as on other tissue-specific stem cells, such as renal, prostate, and corneal stem cells. The putative murine homologue, prominin, which is expressed on neuroepithelial and epithelial mouse cells, was dentified.

Alternative names

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

Detailed product information

Technical specifications

Clone293C3
Clonalitymonoclonal
Isotypemouse IgG2bκ
Isotype controlIsotype Control Antibody, mouse IgG2b
Hostmouse
Type of antibodyPrimary antibodies
Specieshuman
AntigenCD133/2
Alternative names of antigenPROM1, AC133, CORD12, MCDR2, MSTP061, PROML1, RP41, STGD4
Molecular mass of antigen [kDa]117
Distribution of antigenendothelial cells, epithelial cells, red blood cells, hematopoietic stem and progenitor cells, ES and iPS cells, brain, heart, kidney, liver, lung, pancreas, placenta
Entrez Gene ID8842
RRIDAB_2726014, AB_2726287, AB_2726012, AB_2726285, AB_2726010, AB_2726288, AB_2726013, AB_2726286, AB_2726011, AB_2801731, AB_2801730, AB_244343, AB_2726289

Resources for CD133/2 Antibody, anti-human

Reviews for CD133/2 Antibody, anti-human

Human Antibody CD133-APC

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CD133/2 (293C3)-APC, human (130-113-184)

For FACS sorting or analysis of the CD133 epitope, single cells were stained for 30 min at 4°C with monoclonal antibody CD133-APC (CD133/2; Miltenyi Biotec, 10 µl per test up to 10^7 cells in a total volume of 100 µL buffer). Viable CD133+/CD133- populations were sorted with isotype control -based gating parameters.

References for CD133/2 Antibody, anti-human

Publications

  1. Miraglia, S. et al. (1997) A novel five-transmembrane hematopoietic stem cell antigen: isolation, characterization, and molecular cloning. Blood 90(12): 5013-5021
  2. Yin, A. H. et al. (1997) AC133, a novel marker for human hematopoietic stem and progenitor cells. Blood 90: 5002-5012
  3. 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
  4. 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
  5. Uchida, N. et al. (2000) Direct isolation of human central nervous system stem cells. Proc. Natl. Acad. Sci. U.S.A. 97: 14720-14725
  6. 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
  7. Pfenninger, C. V. et al. (2007) CD133 is not present on neurogenic astrocytes in the adult subventricular zone, but on embryonic neural stem cells, ependymal cells, and glioblastoma cells. Cancer Res. 67(12): 5727-5736
  8. Bussolati, B. et al. (2005) Isolation of renal progenitor cells from adult human kidney. Am. J. Pathol. 166: 545-555
  9. Bühring, H. J. et al. (1999) Expression of novel surface antigens on early hematopoietic cells. Ann. N. Y. Acad. Sci. 872: 25-39
  10. Richardson, G. et al. (2004) CD133, a novel marker for human prostatic epithelial stem cells. J. Cell. Sci. 117: 3539-3545
  11. 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
  12. 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
  13. Bühring, H. J. et al. (1991) The product of the proto-oncogene c-kit (P145c-kit) is a human bone marrow surface antigen of hematopoietic precursor cells which is expressed on a subset of acute non-lymphoblastic cells. Leukemia 5: 854-860
  14. Kurian, L. et al. (2013) Conversion of human fibroblasts to angioblast-like progenitor cells. Nat. Methods 10(1): 77-83
  15. Cox, J. L. et al. (2012) Elevating SOX2 levels deleteriously affects the growth of medulloblastoma and glioblastoma cells. PLoS One 7(8): e44087
  16. Donnenberg, A. D. et al. (2013) KIT (CD117) expression in a subset of non-small cell lung carcinoma (NSCLC) patients. PLoS One 7(12): e52885
  17. 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
  18. 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
  19. Kang, T. W. et al. (2014) Growth arrest and forced differentiation of human primary glioblastoma multiforme by a novel small molecule. Sci Rep 4: 5546
  20. Hawley, R. G. et al. (2006) Hematopoietic stem cells. Meth. Enzymol. 419: 149-179
  21. Golden, J. E. et al. (2012) Human umbilical cord blood cells alter blood and spleen cell populations after stroke. Transl Stroke Res 3(4): 491-499
  22. Wang, L. et al. (2013) Enrichment of prostate cancer stem-like cells from human prostate cancer cell lines by culture in serum-free medium and chemoradiotherapy. Int. J. Biol. Sci. 9(5): 472-479

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