Cell Signaling Technology

Product Pathways - Development

StemLight™ Pluripotency Surface Marker Antibody Kit #9094

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
SSEA4 (MC813) Mouse mAb #4755 40 µl IF-IC F H N/A Mouse IgG3
TRA-1-60(S) (TRA-1-60(S)) Mouse mAb #4746 40 µl W IHC-P IF-IC F H 200-400 Mouse IgM
TRA-1-81 (TRA-1-81) Mouse mAb #4745 40 µl IHC-P IF-IC F H 200-400 Mouse IgM

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

TRA-1-60(S) (TRA-1-60(S)) Mouse mAb detects endogenous levels of the neuraminidase-sensitive TRA-1-60 antigen. TRA-1-81 (TRA-1-81) Mouse mAb detects endogenous levels of TRA-1-81 antigen. SSEA4 (MC813) Mouse mAb detects endogenous levels of SSEA4 antigen.

IF-IC

IF-IC

Confocal immunofluorescent analysis of NTERA-2 cells, using TRA-1-60(S) (TRA-1-60(S)) Mouse mAb, TRA-1-81 (TRA-1-81) Mouse mAb, and SSEA4 (MC813) Mouse mAb. Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Description

The StemLight™ Surface Marker Kit contains a panel of antibodies for the detection of antigens that are specifically expressed on the surface of 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.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with human embryonic carcinoma 2102Ep cl.2A6 cells.

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.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 (1). TRA-1-60(S) and TRA-1-81 antibodies recognize different proteoglycan epitopes on variants of the same protein, podocalyxin (2). These epitopes are neuraminidase 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 (3).

  1. Henderson, J.K. et al. (2002) Stem Cells 20, 329-37.
  2. Draper, J.S. et al. (2002) J Anat 200, 249-58.
  3. Schopperle, W.M. and DeWolf, W.C. (2007) Stem Cells 25, 723-30.

Application References

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Protocols

* Product-specific protocol.

For Research Use Only. Not For Use In Diagnostic Procedures.

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