Revision 4

#9656

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Kit Includes

Quantity

Applications

Dilution

Isotype

Oct-4A (C30A3) Rabbit mAb #94310

100 tests

IF-IC

1:200

Rabbit IgG

Sox2 (D6D9) Rabbit mAb #34516

100 tests

IF-IC

1:200

Rabbit IgG

Nanog (D73G4) XP® Rabbit mAb #99399

100 tests

IF-IC

1:200

Rabbit IgG

SSEA4 (MC813) Mouse mAb #43782

100 tests

IF-IC

1:200

Mouse IgG3

TRA-1-60(S) (TRA-1-60(S)) Mouse mAb #61220

100 tests

IF-IC

1:200

Mouse IgM

TRA-1-81 (TRA-1-81) Mouse mAb #83321

100 tests

IF-IC

1:200

Mouse IgM

Applications Key: IF-IC=Immunofluorescence (Immunocytochemistry)

Description

The StemLight™ Pluripotency IF Antibody Sampler 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.


Kit Usage Information

Protocols
34516: Immunofluorescence
94310: Immunofluorescence
99399: Immunofluorescence
43782: Immunofluorescence
61220: Immunofluorescence
83321: Immunofluorescence

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).

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.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

Cell Signaling Technology Logo
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.
undefined Image 1: StemLight<sup>™</sup> Pluripotency IF Antibody Sampler Kit
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).
undefined Image 2: StemLight<sup>™</sup> Pluripotency IF Antibody Sampler Kit
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).
undefined Image 3: StemLight<sup>™</sup> Pluripotency IF Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

Cell Signaling Technology Logo
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.
undefined Image 4: StemLight<sup>™</sup> Pluripotency IF Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.