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SCR001 | ES Cell Characterization Kit

SCR001
100 assays  
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      Overview

      Replacement Information

      Key Specifications Table

      Key Applications
      ICC
      Description
      Catalogue NumberSCR001
      Brand Family Chemicon®
      Trade Name
      • Chemicon
      DescriptionES Cell Characterization Kit
      OverviewThe ES Cell Characterization Kit consists of two components used for alkaline phosphatase activity determination as well as four ES cell-specific antibodies required to perform 100 tests (including controls).

      INTRODUCTION

      Stem cells have become the subject of extensive investigation recently, partly due to their therapeutic potential and because they raise several fundamental issues concerning the regulation of proliferation and differentiation.

      Embryonic stem (ES) cells are derived from totipotent cells of the early mammalian embryo and are capable of unlimited, undifferentiated proliferation in vitro [Evans and Kaufman, 1981; Martin, 1981; Morrison, 1997]. Undifferentiated mouse ES cells can be maintained for an extensive period of time in media containing the cytokine, leukemia-inhibitory factor (LIF) or CHEMICON's proprietary ES cell culture reagent, ESGRO® [Smith, 1988; Williams, 1988]. However, upon removal of LIF from the culture medium, in vitro, the mouse ES cells start to differentiate into cells derived from all three germ layers. In contrast, human ES cultures require mouse fibroblasts as feeder cells and cannot be maintained with LIF for self-renewal [Thomson and Marshall, 1998; Shamblott, 1988].

      The undifferentiated state of the embryonic stem cell is characterized by a high level of expression of alkaline phosphatase (AP) [Pease, 1990] and the stem cell transcription factor, Oct-4. Nevertheless, these ES cells also exhibit marked differences from their murine counterparts in regards to their expression of stage-specific embryonic antigen (SSEA), that typify undifferentiated human ES and embryonic carcinoma (EC) cells.

      SSEA-1, a carbohydrate antigen, is a fucosylated derivative of type 2 polylactosamine and appears during late cleavage stages of mouse embryos. It is strongly expressed by undifferentiated, murine ES cells [Solter and Knowles, 1978; Gooi, 1981]. Upon differentiation, murine ES cells are characterized by the loss of SSEA-1 expression and may be accompanied, in some instances, by the appearance of SSEA-3 and SSEA-4 [Solter, 1979]. In contrast, human ES and EC cells typically express SSEA-3 and SSEA-4 but not SSEA-1, while their differentiation is characterized by downregulation of SSEA-3 and SSEA-4 and an upregulation of SSEA-1 [Andrews, 1984; Fenderson and Andrews; 1987]. Undifferentiated, human ES cells also express the keratin sulphate-associated antigens, TRA-1-60 and TRA-1-81 [Andrews and Banting, 1984].

      CHEMICON's ES Cell Characterization Kit (Catalog number SCR001) is a specific and sensitive tool for the phenotypic assessment of the differentiation status of ES cells by measuring their AP activity, cell-surface stage-specific antigens (SSEA-1, SSEA-4) as well as expression of TRA-1-60, TRA-1-81 antigens.
      Materials Required but Not Delivered1. Fixative

      2. 1 x Rinse Buffer (e.g. TBST: 20mM Tris-HCl, pH 7.4, 0.15M NaCl, 0.05% Tween-20)

      3. Blocking solution

      4. Fluorescent-labeled secondary antibodies

      5. Hematoxylin

      6. Microscope
      References
      Product Information
      Components
      • Fast Red Violet solution (0.8g/L stock) (Part No. 90239). One 15mL bottle.
      • Napthol AS-BI phosphate solution (4mg/mL) in AMPD buffer (2mol/L), pH 9.5 (Part No. 90234). One 15mL bottle.
      • MS X SSEA-1, IgM, clone MC-480 (Part No. 90230). One vial containing 100μl of 1mg/mL monoclonal antibody.
      • MS X SSEA-4, IgG, clone MC-813-70 (Part No. 90231). One vial containing 100μl of 1mg/mL monoclonal antibody.
      • MS X TRA-1-60, IgM, clone TRA-1-60 (Part No. 90232). One vial containing 100μl of 1mg/mL monoclonal antibody
      • MS X TRA-1-81, IgM, clone TRA-1-81 (Part No: 90233). One vial containing 100μl of 1mg/mL monoclonal antibody.
      Applications
      ApplicationThe Embryonic Stem Cell Characterization Kit phenotypically assesses the differentiation status of ES cells by measuring their AP activity, cell-surface stage-specific antigens (SSEA-1, SSEA-4) as well as expression of TRA-1-60, TRA-1-81 antigens.
      Key Applications
      • Immunocytochemistry
      Biological Information
      Species Reactivity
      • Human
      • Mouse
      • Rat
      Stem Cell Type
      • Human Embryonic Stem Cells
      • Mouse Embryonic Stem Cells
      • Induced Pluripotent Stem Cells
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsWhen stored at 2-8º C, the kit components are stable up to the expiration date. Do not freeze or expose to elevated temperatures. Discard any remaining reagents after the expiration date.
      Packaging Information
      Material Size100 assays
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      SDS

      Title

      Safety Data Sheet (SDS) 

      Certificates of Analysis

      TitleLot Number
      ES Cell Characterization Kit -2550570 2550570

      References

      Reference overviewPub Med ID
      Reprogrammed keratinocytes from elderly type 2 diabetes patients suppress senescence genes to acquire induced pluripotency.
      Ohmine, S; Squillace, KA; Hartjes, KA; Deeds, MC; Armstrong, AS; Thatava, T; Sakuma, T; Terzic, A; Kudva, Y; Ikeda, Y
      Aging 4 60-73 2012

      Show Abstract
      22308265 22308265
      Generation of transgene-free lung disease-specific human induced pluripotent stem cells using a single excisable lentiviral stem cell cassette.
      Aba Somers,Jyh-Chang Jean,Cesar A Sommer,Amel Omari,Christopher C Ford,Jason A Mills,Lei Ying,Andreia Gianotti Sommer,Jenny M Jean,Brenden W Smith,Robert Lafyatis,Marie-France Demierre,Daniel J Weiss,Deborah L French,Paul Gadue,George J Murphy,Gustavo Mostoslavsky,Darrell N Kotton
      Stem cells (Dayton, Ohio) 28 2010

      Show Abstract
      20715179 20715179
      Cardiac commitment of primate embryonic stem cells.
      Leschik J. et al.
      Nat. Protoc. 3(9) 1381-1387 2008

      Show Abstract
      18772864 18772864
      Co-localization of angiotensin-converting enzyme 2-, octomer-4- and CD34-positive cells in rabbit atherosclerotic plaques.
      Anthony Zulli, Sudarshan Rai, Brian F Buxton, Louise M Burrell, David L Hare
      Experimental physiology 93 564-9 2008

      Show Abstract
      18192339 18192339
      Establishment and characterization of new human embryonic stem cell lines.
      Findikli, Necati, et al.
      Reprod. Biomed. Online, 10: 617-27 (2005) 2005

      Show Abstract
      15949219 15949219

      Brochure

      Title
      Human Stem Cell Systems
      Murine Embryonic Stem Cell Culture

      Data Sheet

      Title
      ES Cell Characterization Kit - Data Sheet