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ES Cell Marker Sample Kit

Description:
ES Cell Marker Sample Kit
Trade Name:
Chemicon (Millipore)
Qty/Pk:
1 kit
Product Overview:
The ES Cell Marker Sample Kit includes six monoclonal antibodies for the analysis of ES cell marker expression.

Introduction:

Embryonic stem (ES) cells are derived from totipotent cells of the early mammalian embryo and are capable of unlimited, undifferentiated proliferation in vitro (Evans & Kaufman, 1981; Martin, 1981; Morrison SJ et al., 1981). Undifferentiated mouse ES cells can be maintained for extended periods in media containing the cytokine, Leukemia Inhibitory Factor (LIF) or CHEMICON®'s proprietary ES Cell culture reagent ESGROâ (Smith AG et al., 1988; Williams, RL 1988). Upon removal of LIF from the culture medium, in vitro, the mouse ES cells 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 JA & Marshall VS 1998; Shamblott MJ et al., 1998).

The undifferentiated state of the ES cells is characterized by high level of expression of the POU transcription factor Octamer-4 (Oct-4) (Pesce M et al., 1998). This relationship between Oct-4 and pluripotency has seen this transcription factor emerge as a marker of pluripotent stem cells. Undifferentiated human and murine pluripotent Embryonic Stem (ES) and Embryonic Carcinoma (EC) cells express Oct-4. Additionally, murine Embryonic Germ (EG) cells are also known to express Oct-4. Following stem cell differentiation, the level of Oct-4 expression decreases.

Pluripotent stem cells can also be characterized by the expression of a number of cell surface antigens. 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 D, 1978; Gooi HC, 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 D, 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 down regulation of SSEA-3 and SSEA-4 and an up regulation of SSEA-1 (Andrews PW, 1984; Fenderson BA, Andrews PW et al., 1987). Undifferentiated, human ES cells also express the keratan sulphate-associated antigens, TRA-1-60 and TRA-1-81 (Andrews PW, Banting GS et al., 1984).

CHEMICON®'s ES Cell Marker Sample Kit (Catalog number SCR002) contains monoclonal antibodies for the detection of the cell-surface Stage-Specific Embryonic Antigens (SSEA-1, SSEA-3 & SSEA-4), expression of TRA-1-60, TRA-1-81 antigens and Oct-4. This kit permits the phenotypic analysis of the differentiation status of ES cells by determining stem cell marker expression.
Applications:
CHEMICON's Embryonic Stem (ES) Cell Marker Sample Kit contains monoclonal antibodies for the detection of the cell-surface Stage-Specific Embryonic Antigens (SSEA-1, SSEA-3 & SSEA-4), expression of TRA-1-60, TRA-1-81 antigens & Oct-4.
Key Applications:
Immunocytochemistry
Species Reactivity:
  • Human
  • Mouse
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.
Brand Family:
Chemicon
Storage Conditions:
When stored at 2-8°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.
Product Name:
ES Cell Marker Sample Kit
Materials Required but Not Delivered:
1. Fixative Solution (4% paraformaldehyde / PBS)

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

3. Blocking solution (e.g. 4% normal goat serum / PBS)

4. Microscope

5. Fluorochrome conjugated secondary antibodies
Components:
  • MS X SSEA-1, IgM, clone MC-480: (Part No. MAB4301-20). One vial containing 20μL of 1mg/mL monoclonal antibody.
  • RT X SSEA-3, IgM, clone MC-631: (Part No. MAB4303-20). One vial containing 20μL of 1 mg/mL monoclonal antibody.
  • MS X SSEA-4, IgG, clone MC-813-70: (Part No. MAB4304-20). One vial containing 20μL of 1mg/mL monoclonal antibody.
  • MS X TRA-1-60, IgM, clone TRA-1-60: (Part No. MAB4360-20). One vial containing 20μL of 1mg/mL monoclonal antibody
  • MS X TRA-1-81, IgM, clone TRA-1-81: (Part No: MAB4381-20). One vial containing 20μL of 1mg/mL monoclonal antibody.
  • MS X Oct-4, IgG, clone 9E3.2: (Part number: MAB4305-20). One vial containing 20μL of 1mg/mL monoclonal antibody.
Packaging:
Sufficient for 20 reactions (including controls).

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