Revision 2

#9656Store at -20C

1 Kit

(100 tests)

Cell Signaling Technology

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Web: info@cellsignal.com cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Kit Includes Quantity Applications Dilution Isotype
Oct-4A (C30A3) Rabbit mAb 100 tests IF-IC 1:200 Rabbit IgG
Sox2 (D6D9) Rabbit mAb 100 tests IF-IC 1:200 Rabbit IgG
Nanog (D73G4) XP® Rabbit mAb 100 tests IF-IC 1:200 Rabbit IgG
SSEA4 (MC813) Mouse mAb 100 tests IF-IC 1:200 Mouse IgG3
TRA-1-60(S) (TRA-1-60(S)) Mouse mAb 100 tests IF-IC 1:200 Mouse IgM
TRA-1-81 (TRA-1-81) Mouse mAb 100 tests IF-IC 1:200 Mouse IgM

Applications Key: IF-IC=Immunofluorescence (Immunocytochemistry)

Description

The StemLight® Pluripotency Antibody Kit contains a panel of antibodies for the detection of proteins that are specifically expressed in human pluripotent cells. The kit can be used to track the pluripotent potential of human embryonic stem (ES) or induced pluripotent (iPS) cells. The loss of these markers indicates a loss of pluripotency or differentiation of the culture. The kit components are pre-optimized for parallel use in immunofluorescent analysis. Enough reagents are provided for 100 assays based on a working volume of 100 µl.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

Pluripotency is the ability of a cell to differentiate into cell types of the three germ layers, the endoderm, ectoderm and mesoderm. It is a property shared by embryonic stem cells, embryonic carcinoma and induced pluripotent cells.
Oct-4, Sox2 and Nanog are key transcriptional regulators that are highly expressed in pluripotent cells (1). Together they form a transcriptional network that maintains cells in a pluripotent state (2,3). Over-expression of Oct-4 and Sox2 along with Klf4 and c- Myc can induce pluripotency in both mouse and human somatic cells, highlighting their roles as key regulators of the transcrip- tional network necessary for renewal and pluripotency (4-5). It has also been demonstrated that overexpression of Oct-4, Sox2, Nanog and Lin28 can induce pluripotency in human somatic cells (6). Upon differentiation of pluripotent cultures, expression of Oct-4, Nanog and Sox2 is downregulated.
SSEA4, TRA-1-81 and TRA-1-60 antibodies recognize antigens expressed on the cell surface of all pluripotent cells. SSEA4 recognizes a glycolipid carbohydrate epitope (7). TRA-1-60(S) and TRA-1-81 antibodies recognize different proteoglycan epitopes on variants of the same protein, podocalyxin (8). These epitopes are neuraminadase sensitive and resistant, respectively. Reactivity of SSEA4, TRA-1-81 and TRA-1-60 antibodies with their respective cell surface markers are lost upon differentiation of pluripotent cells, corresponding with a loss of pluripotent potential (9).

  1. Looijenga, L.H. et al. (2003) Cancer Res 63, 2244-50.
  2. Pesce, M. and Schöler, H.R. (2001) Stem Cells 19, 271-8.
  3. Pan, G. and Thomson, J.A. (2007) Cell Res 17, 42-9.
  4. Takahashi, K. and Yamanaka, S. (2006) Cell 126, 663-76.
  5. Okita, K. et al. (2007) Nature 448, 313-7.
  6. Yu, J. et al. (2007) Science 318, 1917-20.
  7. Henderson, J.K. et al. (2002) Stem Cells 20, 329-37.
  8. Draper, J.S. et al. (2002) J Anat 200, 249-58.
  9. Schopperle, W.M. and DeWolf, W.C. (2007) Stem Cells 25, 723-30.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    StemLight® is a registered trademark of Cell Signaling Technology, Inc.
    U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    Revision 2
    #9656

    StemLight Pluripotency Antibody Kit

    StemLight™ Pluripotency Antibody Kit: Image 1 Expand Image
    Confocal immunofluorescent analysis of NTERA-2 cells, untreated (left panel) or retinoic acid-treated (10 µM all-trans RA for 5 days) (right panel), using Neurofilament-L (C28E10) Rabbit mAb #2837 (green, upper), and β3-Tubulin (TU-20) Mouse mAb #4466 (green, lower). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). Note the appearance of neuronal markers and structures as cells differentiate along the neuronal lineage with retinoic acid treatment.
    StemLight™ Pluripotency Antibody Kit: Image 2 Expand Image
    Confocal immunofluorescent analysis of NTERA-2 (left) and HeLa cells (right) using Oct-4A (C30A3) Rabbit mAb (green, upper), Sox2 (D6D9) XP® Rabbit mAb (green, middle) and Nanog (D73G4) XP® Rabbit mAb (green, lower).
    StemLight™ Pluripotency Antibody Kit: Image 3 Expand Image
    Projected confocal z-stack of human iPS cells using TRA-1-60(S) (TRA-1-60(S)) Mouse mAb (green, upper left), TRA-1-81 (TRA-1-81) Mouse mAb (green, upper middle), SSEA4 (MC813) Mouse mAb (green, upper right), Oct-4A (C30A3) Rabbit mAb (green, lower left), Sox2 (D6D9) XP® Rabbit mAb (green, lower middle) and Nanog (D73G4) XP® Rabbit mAb (green, lower right). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    StemLight™ Pluripotency Antibody Kit: Image 4 Expand Image
    Confocal immunofluorescent analysis of NTERA-2 cells, untreated (left panel) or retinoic acid-treated (10 µM all-trans RA for 5 days) (right panel), using TRA-1-60(S) (TRA-1-60(S)) Mouse mAb (green, upper), TRA-1-81 (TRA-1-81) Mouse mAb (green, middle) and SSEA4 (MC813) Mouse mAb (green, lower). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). Note the loss of pluripotency markers (green) as cells differentiate along the neuronal lineage with retinoic acid treatment.