CD133 (Prominin-1) Monoclonal Antibody (13A4), APC, ebioscience Catalog Number Product data sheet

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1 Website: thermofisher.com/ebioscience Customer Service (US): thermofisher.com/contactus CD133 (Prominin-1) Monoclonal Antibody (13A4), APC, ebioscience Catalog Number Product data sheet Details Size 50 µg Host/Isotope Class Type Clone Conjugate Form Concentration Purification Storage buffer Contains Storage Conditions Rat / IgG1, kappa Monoclonal Antibody 13A4 APC Liquid 0.2 mg/ml Affinity chromatography PBS, ph 7.2, with 0.1% gelatin 0.09% sodium azide 4 C, store in dark, DO NOT FREEZE! Species Reactivity Tested species reactivity Dog, Mouse Published species reactivity Mouse, Human, Not Applicable Tested Applications Dilution * Flow Cytometry (Flow) µg/test Published Applications Flow Cytometry (Flow) See 8 publications below Immunofluorescence (IF) See 2 publications below Miscellaneous PubMed (MISC) See 1 publications below * Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls. Product specific information Description: The 13A4 monoclonal antibody recognizes mouse Prominin-1 (sometimes also referred to as CD133 and, in the case of the human orthologue, as AC133), a kda pentaspan transmembrane (5-TM) domain glycoprotein. Prominin-1 is expressed on primitive cells such as hematopoietic stem and progenitor cells, neural and endothelial stem cells, retina and retinoblastoma, as well as developing epithelium. To date, the function and ligand of Prominin-1 are unknown. The 13A4 antibody does not cross react with rat, human, chicken, or Drosophila antigen but has been reported to work in canine /dog. Applications Reported: The 13A4 antibody has been reported for use in flow cytometric analysis. Applications Tested: This 13A4 antibody has been tested by flow cytometric analysis of mouse bone marrow cells. This can be used at less than or equal to µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µl. Cell number should be determined empirically but can range from 10^5 to 10^8 cells /test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Excitation: nm; Emission: 660 nm; Laser: Red Laser. Filtration: 0.2 µm post-manufacturing filtered. Background/Target Information The CD133 (also named AC133) antigen is a 97 kda glycoprotein, with 5 transmembrane domains, that shows up as a 120 kda band on protein gels due to glycosylation. It is a cell surface marker expressed by immature hematopoietic stem cells but not mature blood cells. In addition to hematopoietic stem and progenitor cells, CD133 has been found to be a marker for other stem and progenitor cells including neural and embryonic stem cells. It has been shown to be expressed in cancers, including some leukemias and brain tumours, that may be derived from stem cells. Low level expression could also be detected in the kidney, pancreas, placenta, and fetal liver.

2 Product Images For CD133 (Prominin-1) Monoclonal Antibody (13A4), APC, ebioscience CD133 (Prominin-1) Antibody ( ) Staining of BALB/c bone marrow cells with Anti-Mouse CD11b FITC (Product # ) and 0.06 µg of Rat IgG1 kappa Isotype Control APC (Product # ) (left) or 0.06 µg of Anti-Mouse CD133 (Prominin-1) APC (right). Total viable cells were used for analysis. CD133 (Prominin-1) Antibody in Flow Cytometry (Flow) 6 ED Figure 6. Defects in mammary glands in DS mice models a,b. mrna quantification of Usp16 and different Hox genes in CD49 high CD24 med mammary cells. Hox1, Hox3 and Hox5 are expressed at higher levels in Ts65Dn cells. c, Representative FACS plot of mammary cells gated on live cells (first row) or live Lin - cells. We observed a perturbation in the overall FACS profile with reduction of basal and luminal cells (indicated gates) in Ts65Dn mice but not in Ts1Cje mice. These experiments were repeated 5 times for each group. d, Quantification of overlap between staining for the basal cytokeratin CK14 (red) and the luminal cytokeratin CK8 (green). Pearson's correlation analyses (Lumosity software) showed a marked increase in cells that co-stain for both cytokeratins in Ts65Dn mammary epithelium. Each experiment was repeated with 3 mice per group. e, infection partially in vivo Usp16 by shrna lentiviral Downregulation of by Ts65Dn rescues the defects shown mammary (p=0.03). Three independent On transplantation experiments were performed. cells d filled by GFP outgrowths is significantly higher upon the right, the percentage of fat pa downregulation of Usp16 (p=0.007). CD133 (Prominin-1) Antibody in Flow Cytometry (Flow)

3 CD133 (Prominin-1) Antibody in Immunofluorescence (IF) Published figure using CD133 (Prominin-1) monoclonal antibody (Product # ) in Immunofluorescence

4 PubMed References For CD133 (Prominin-1) Monoclonal Antibody (13A4), APC, ebioscience 8 Flow Cytometry References was used in Flow cytometry/cell sorting to investigate the precise role and expression pattern of brain-expressed X- linked genes in endocrine and tissue stem/progenitor cells in mice. Human / 1:33 Mouse / 1:5 The Journal of biological chemistry (Oct 2014; 289: 29892) "Gene targeting study reveals unexpected expression of brain-expressed X-linked 2 in endocrine and tissue stem /progenitor cells in mice." Author(s):Ito K,Yamazaki S,Yamamoto R,Tajima Y,Yanagida A,Kobayashi T,Kato-Itoh M,Kakuta S,Iwakura Y,Nakauchi H, Kamiya A PubMed Article URL: was used in Flow cytometry/cell sorting to develop a validated protocol enabling the co-detection and co-expression of surface molecules on neural cell populations. PloS one (Feb 2014; 8: null) "Combined flow cytometric analysis of surface and intracellular antigens reveals surface molecule markers of human neuropoiesis." Author(s):Turaç G,Hindley CJ,Thomas R,Davis JA,Deleidi M,Gasser T,Karaöz E,Pruszak J PubMed Article URL: was used in Flow cytometry/cell sorting to compare the tumorigenic potential and cell fate of cells with self-renewing potential in an in vivo melanoma model. PloS one (Nov 2013; 8: null) "Association between tumorigenic potential and the fate of cancer cells in a syngeneic melanoma model." Author(s):Krelin Y,Berkovich L,Amit M,Gil Z PubMed Article URL: Nature (Sep 2013; 501: 380) "Usp16 contributes to somatic stem-cell defects in Down's syndrome." Author(s):Adorno M,Sikandar S,Mitra SS,Kuo A,Nicolis Di Robilant B,Haro-Acosta V,Ouadah Y,Quarta M,Rodriguez J,Qian D, Reddy VM,Cheshier S,Garner CC,Clarke MF PubMed Article URL: was used in Flow cytometry/cell sorting to investigate the presence of cardiac-resident progenitor cells with high aldehyde dehydrogenase activity isolated from mouse hearts. BioMed research international (Sep 2013; 2013: null) "Characterization of cardiac-resident progenitor cells expressing high aldehyde dehydrogenase activity." Author(s):Roehrich ME,Spicher A,Milano G,Vassalli G PubMed Article URL: was used in Flow cytometry/cell sorting to develop an optimised method for studying tumour angiogenesis. Angiogenesis (Sep 2010; 13: 251) "Minimally invasive assessment of tumor angiogenesis by fine needle aspiration and flow cytometry." Author(s):Sottnik JL,Guth AM,Mitchell LA,Dow SW PubMed Article URL: was used in Flow cytometry/cell sorting to assess the assumption that CD133 is expressed constitutively in neural precursor cells. PloS one (Sep 2009; 4: null) "CD133 (Prominin) negative human neural stem cells are clonogenic and tripotent." Author(s):Sun Y,Kong W,Falk A,Hu J,Zhou L,Pollard S,Smith A PubMed Article URL: 2 Immunofluorescence References Proceedings of the National Academy of Sciences of the United States of America (Jan 2007; 104: 175) "Conserved markers of fetal pancreatic epithelium permit prospective isolation of islet progenitor cells by FACS." Author(s):Sugiyama T,Rodriguez RT,McLean GW,Kim SK PubMed Article URL:

5 Cancer research (Jun 2007; 67: 5727) "CD133 is not present on neurogenic astrocytes in the adult subventricular zone, but on embryonic neural stem cells, ependymal cells, and glioblastoma cells." Author(s):Pfenninger CV,Roschupkina T,Hertwig F,Kottwitz D,Englund E,Bengzon J,Jacobsen SE,Nuber UA PubMed Article URL: 1 Miscellaneous PubMed References Proceedings of the National Academy of Sciences of the United States of America (Jan 2007; 104: 175) "Conserved markers of fetal pancreatic epithelium permit prospective isolation of islet progenitor cells by FACS." Author(s):Sugiyama T,Rodriguez RT,McLean GW,Kim SK PubMed Article URL: Cancer research (Jun 2007; 67: 5727) "CD133 is not present on neurogenic astrocytes in the adult subventricular zone, but on embryonic neural stem cells, ependymal cells, and glioblastoma cells." Author(s):Pfenninger CV,Roschupkina T,Hertwig F,Kottwitz D,Englund E,Bengzon J,Jacobsen SE,Nuber UA PubMed Article URL: