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9092
StemLight™ iPS Cell Reprogramming Antibody Kit
Primary Antibodies
Antibody Sampler Kit

StemLight™ iPS Cell Reprogramming Antibody Kit #9092

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StemLight™ iPS Cell Reprogramming Antibody Kit: Image 1

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).

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 2

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).

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 3

Flow cytometric analysis of HeLa cells (blue) and NCCIT cells (green) using Oct-4A (C30A3) Rabbit mAb.

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 4

Flow cytometric analysis of HeLa cells (blue) and NTERA2 cells (green) using Sox2 (D6D9) XP® Rabbit mAb.

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 5

Flow cytometric analysis of HeLa cells (blue) and NTERA-2 cells (green) using Nanog (D73G4) XP® Rabbit mAb.

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 6

Flow cytometric analysis of HeLa cells (blue) and NTERA2 cells (green) using LIN28A (D84C11) XP® Rabbit mAb.

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 7

Immunohistochemical analysis of paraffin-embedded human seminoma using LIN28A (D84C11) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 8

Western blot analysis of extracts from various cell lines using KLF4 antibody.

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 9

Flow cytometric analysis of Raji cells using c-Myc (D84C12) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 10

Confocal immunofluorescent analysis of NTERA2 (left) and mouse embryonic stem (mES) cells growing on mouse embryonic fibroblast (MEF) feeder cells (right) using Oct-4A (C30A3) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 11

Confocal immunofluorescent analysis of NTERA2 (left) and HeLa (right) cells using Sox2 (D6D9) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 12

Confocal immunofluorescent analysis of NTERA-2 cells (left) and HeLa cells (right) using Nanog (D73G4) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 13

Confocal immunofluorescent analysis of NTERA-2 cells using Lin28A (D84C11) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 14

Immunohistochemical analysis of paraffin-embedded human tumor tissues: seminoma (top-left), prostate carcinoma (top-right), neuroendocrine lung carcinoma (bottom-left) and ductal breast carcinoma (bottom-right) using LIN28A (D84C11) XP® Rabbit mAb.

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 15

Confocal immunofluorescent analysis of HeLa cells, mock-transfected (left) or transfected with SignalSilence® c-Myc siRNA I #6341 (right), using c-Myc (D84C12) Rabbit mAb (green). Actin filaments have been labeled wth DY-554 phalloidin (red).

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 16

Western blot analysis of extracts from NTERA2 and mouse embryonic stem cells (mESCs) using Oct-4A (C30A3) Rabbit mAb.

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 17

Immunohistochemical analysis of paraffin-embedded human seminoma using Nanog (D73G4) XP® Rabbit mAb.

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 18

Immunohistochemical analysis of paraffin-embedded IGROV-1 cell pellet (left, positive) or HCT 116 cell pellet (right, negative) using LIN28A (D84C11) XP® Rabbit mAb.

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 19

Western blot analysis of extracts from control HEK293 cells (lane 1) or c-Myc knockout HEK293 cells (lane 2) using c-Myc (D84C12) Rabbit mAb Antibody, #5605 (upper) or β-actin (13E5) Rabbit mAb, #4970 (lower). The absence of signal in the c-Myc knockout HEK293 cells confirms specificity of the antibody for c-Myc.

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 20

Western blot analysis of extracts from NCCIT, NTERA-2 and iPS cells using Nanog (D73G4) XP® Rabbit mAb.

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 21

Immunohistochemical analysis of paraffin-embedded normal human tissues: testis (top-left), prostate (top-right), pancreas (bottom-left) and kidney (bottom-right) using LIN28A (D84C11) XP® Rabbit mAb.

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 22

Western blot analysis of extracts from HeLa cells, mock transfected or transfected with SignalSilence® c-Myc siRNA I #6341, using c-Myc (D84C12) Rabbit mAb.

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 23

Western blot analysis of extracts from NTERA2 and NCCIT cells using Sox2 (D6D9) XP® Rabbit mAb.

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 24

Western blot analysis of extracts from various cell lines using c-Myc (D84C12) Rabbit mAb.

StemLight™ iPS Cell Reprogramming Antibody Kit: Image 25

Western blot analysis of extracts of NTERA-2 and IGROV1 cells using LIN28A (D84C11) XP® Rabbit mAb.

To Purchase # 9092T
Product # Size Price
9092T
1 Kit  (6 x 20 µl) $ 445

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Oct-4A (C30A3) Rabbit mAb 2840 20 µl
  • WB
  • IF
  • F
H M 45 Rabbit IgG
Sox2 (D6D9) XP® Rabbit mAb 3579 20 µl
  • WB
  • IF
  • F
H 35 Rabbit 
Nanog (D73G4) XP® Rabbit mAb 4903 20 µl
  • WB
  • IHC
  • IF
  • F
H 42 Rabbit IgG
LIN28A (D84C11) XP® Rabbit mAb 3695 20 µl
  • WB
  • IHC
  • IF
  • F
H 26 Rabbit IgG
KLF4 Antibody 4038 20 µl
  • WB
H M Mk 65 Rabbit 
c-Myc (D84C12) Rabbit mAb 5605 20 µl
  • WB
  • IF
  • F
H M R 57-65 Rabbit IgG

Product Description

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) 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.

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.

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's 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's 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.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
StemLight™ is a trademark of Cell Signaling Technology, Inc.
DRAQ5 is a registered trademark of Biostatus Limited.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.