Product Pathways - Stem Cell and Lineage Markers
StemLite™ Pluripotency Kit #9656
| 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 Antibody | 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)
Specificity / Sensitivity
Each antibody in the StemLite™ Pluripotency Kit detects endogenous levels of their respective pluripotency marker proteins.
IF-IC
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 #4746 (green, upper), TRA-1-81 (TRA-1-81) Mouse mAb #4745 (green, middle) and SSEA4 (MC813) Mouse mAb #4755 (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.
IF-IC
Confocal immunofluorescent analysis of NTERA-2 cells, untreated (left panel) or retinoic acid-treated (10 µM all-trans RA for 14 days) (right panel), using Oct-4A (C30A3) Rabbit mAb #2840 (green, upper), Sox2 (D6D9) XP™ Rabbit mAb #3579 (green, middle) and Nanog Antibody #3580 (green, lower). Actin filaments have been labeled with DY-554 phalloidin (red). Note the loss of pluripotency markers (green) as cells differentiate along the neuronal lineage with retinoic acid treatment.
IF-IC
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.
Source / Purification
Nanog Antibody was produced by immunizing animals with a synthetic peptide corresponding to amino acid sequence at the amino terminus of human nanog. Antibodies are purified by Protein A and peptide affinity chromatography. Oct-4A antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of human Oct-4A. Sox2 antibody is produced by immunizing animals with a synthetic peptide corresponding to amino acid sequences at the amino terminus of human Sox2. SSEA4, TRA-1-81, and TRA-1-60(S) antibodies are produced by immunizing animals with human embryonal 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.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 transcriptional network necessary for renewal and pluripotency (4-6). 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.
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
- 2840 Oct-4A (C30A3) Rabbit mAb
- 3579 Sox2 (D6D9) XP™ Rabbit mAb
- 3580 Nanog Antibody
- 4755 SSEA4 (MC813) Mouse mAb
- 4746 TRA-1-60(S) (TRA-1-60(S)) Mouse mAb
- 4745 TRA-1-81 (TRA-1-81) Mouse mAb
- 5605 c-Myc (D84C12) XP™ Rabbit mAb
- 4038 KLF4 Antibody
- 3978 LIN28A (A177) Antibody
- 3695 LIN28A (D84C11) XP™ Rabbit mAb
- 5930 LIN28A (6D1F9) Mouse mAb
- 2837 Neurofilament-L (C28E10) Rabbit mAb
- 4466 β3-Tubulin (TU-20) Mouse mAb
- 4408 Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate)
- 4409 Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 555 Conjugate)
- 4410 Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate)
- 4412 Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate)
- 4413 Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 555 Conjugate)
- 4414 Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate)
- 4084 DRAQ5®
- 4753 Cellular Localization IF Antibody Sampler Kit
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
