Product Pathways - Development
StemLight™ iPS Cell Reprogramming Antibody Kit #9092
|9092S||1 Kit (6 x 40 µl)||---||In Stock||---|
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|Kit Includes||Quantity||Applications||Reactivity||Homology†||MW (kDa)||Isotype|
|Oct-4A (C30A3) Rabbit mAb #2840||40 µl||W, IF-IC, F||H, M||45||Rabbit IgG|
|Sox2 (D6D9) XP® Rabbit mAb #3579||40 µl||W, IHC-P, IF-IC, F||H||Mk, B, Dg, Hr||35||Rabbit|
|Nanog (D73G4) XP® Rabbit mAb #4903||40 µl||W, IHC-P, IF-IC, F||H||Mk||42||Rabbit IgG|
|LIN28A (D84C11) XP® Rabbit mAb #3695||40 µl||W, IF-IC, F||H||R, Mk||26||Rabbit IgG|
|KLF4 Antibody #4038||40 µl||W||H, M, Mk||65||Rabbit|
|c-Myc (D84C12) XP® Rabbit mAb #5605||40 µl||W, IP, IF-IC, F||H, M, R||Mk, Dg, Pg||57-65||Rabbit IgG|
†Species predicted to react based on 100% sequence homology.
Applications Key: W=Western Blotting, IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry, IHC-P=Immunohistochemistry (Paraffin), IP=Immunoprecipitation
Reactivity Key: H=Human, M=Mouse, Mk=Monkey, R=Rat
Confocal immunofluorescent analysis of iPS cells using Oct-4A (C30A3) Rabbit mAb #2840, Sox2 (D6D9) XP® Rabbit mAb #3579, Nanog (D73G4) XP® Rabbit mAb #4903, c-Myc (D84C12) XP® Rabbit mAb #5605, and LIN28A (D84C11) XP® Rabbit mAb #3695 (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Confocal immunofluorescent analysis of lentiviral transduced human fibroblast cells using Oct-4A (C30A3) Rabbit mAb #2840, Sox2 (D6D9) XP® Rabbit mAb #3579, c-Myc (D84C12) XP® Rabbit mAb #5605, and KLF4 Antibody #4038 (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
The StemLight™ iPS cell Reprogramming Antibody Kit contains a panel of antibodies for the detection of various proteins, combinations of which have been used to reprogram somatic cells to Induced Pluripotent Stem (iPS) cells. The kit can be used to track efficiency of expression of the reprogramming factors following transfection, viral transduction and other means of protein delivery. The kit components are pre-optimized for parallel use in immunofluorescent analysis at a standard dilution, but components are also validated for use in other applications --please refer to individual data sheet information for application specific recommendations. Enough reagents are provided for 160 immunofluorescent assays based on a working volume of 100 μl.
Specificity / Sensitivity
Oct-4A (C30A3) Rabbit mAb detects endogenous levels of total Oct-4A protein. Sox2 (D6D9) XP® Rabbit mAb detects endogenous levels of total Sox2 protein. Nanog (D73G4) XP® Rabbit mAb detects endogenous levels of total nanog protein. LIN28A (D84C11) XP® Rabbit mAb detects endogenous levels of total LIN28A protein. c-Myc (D84C12) XP® Rabbit mAb detects endogenous levels of total c-Myc protein. This antibody is not recommended for detection of Myc-tagged fusion proteins (use Cell Signaling Technology cat. #2276 or #2278). KLF4 Antibody detects endogenous levels of total KLF4 protein by western blot, but for immunofluorescence it is only recommended for detection of exogenously expressed KLF4 protein. All components of this kit have been validated for reactivity to human protein, however some components also have additional species cross reactivities. Please see www.cellsignal.com for additional specificity/sensitivity information for individual kit components.
Source / Purification
Monoclonal antibodies are produced by immunizing animals with recombinant protein specific to the amino terminus of human Oct-4A, a synthetic peptide corresponding to residues surrounding Gly179 of human Sox2, a synthetic peptide corresponding to residues near the amino terminus of human nanog protein, a synthetic peptide corresponding to residues near the carboxy terminus of human LIN28A, and a synthetic peptide corresponding
to residues near the amino terminus of c-Myc. Polyclonal antibodies are produced by immunizing animals with synthetic peptide corresponding to residues near the amino terminus of human KLF4. Polyclonal antibodies are purified by Protein A and peptide affinity chromatography.
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 neurominadase 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).
- Looijenga, L.H. et al. (2003) Cancer Res 63, 2244-50.
- Pesce, M. and Schöler, H.R. (2001) Stem Cells 19, 271-8.
- Pan, G. and Thomson, J.A. (2007) Cell Res 17, 42-9.
- Takahashi, K. and Yamanaka, S. (2006) Cell 126, 663-76.
- Okita, K. et al. (2007) Nature 448, 313-7.
- Yu, J. et al. (2007) Science 318, 1917-20.
- Henderson, J.K. et al. (2002) Stem Cells 20, 329-37.
- Draper, J.S. et al. (2002) J Anat 200, 249-58.
- Schopperle, W.M. and DeWolf, W.C. (2007) Stem Cells 25, 723-30.
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For Research Use Only. Not For Use In Diagnostic Procedures.
StemLight™ is a trademark of Cell Signaling Technology, Inc.
DRAQ5® is a registered trademark of Biostatus Limited.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
Select rabbit monoclonal antibodies are developed, validated, and produced at CST using in part technology under license (granting certain rights including those under U.S. Patents No. 5,675,063 and in some instances 7,429,487) from Epitomics, Inc.