Product Pathways - Stem Cell and Lineage Markers
Nanog (D2A3) XP® Rabbit mAb (Mouse Specific) #8822
PhosphoSitePlus® protein, site, and accession data: NANOG
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IP IF-IC F ChIP | M | Endogenous | 29-42 | Rabbit IgG |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
ChIP=Chromatin IP
Reactivity Key:
M=Mouse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
Specificity / Sensitivity
Nanog (D2A3) XP® Rabbit mAb (Mouse Specific) recognizes endogenous levels of total mouse nanog protein. This antibody is expected to recognize nanog 1a and 1b isoforms. This antibody does not cross-react with human nanog.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of mouse nanog protein.
Western Blotting
Western blot analysis of extracts from F9 and murine iPS cells using Nanog (D2A3) XP® Rabbit mAb (Mouse Specific).
Flow Cytometry
Flow cytometric analysis of NIH/3T3 (blue) and F9 (green) cells using Nanog (D2A3) XP® Rabbit mAb (Mouse Specific).
IF-IC
Confocal immunofluorescent analysis of F9 (left) and NIH/3T3 (right) cells using Nanog (D2A3) XP® Rabbit mAb (Mouse Specific) (green). Actin filaments were labeled with DY-554 phalloidin (red).
Chromatin IP
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 F9 cells and either 5 μl of Nanog (D2A3) XP® Rabbit mAb (Mouse Specific) or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Mouse Oct-4 Promoter Primers #4653, SimpleChIP® Mouse XIST Intron 1 Primers #4659, and SimpleChIP® Mouse RPL30 Intron 2 Primers #7015. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Background
Nanog is a homeodomain-containing transcription factor that is essential for the maintenance of pluripotency and self renewal in embryonic stem cells (1). Nanog expression is controlled by a network of factors including Sox2 and the key pluripotency regulator Oct-4 (1). Recent advances in somatic cell reprogramming have utilized viral expression of combinations of transcription factors including nanog, Oct-4, Sox2, KLF4, c-Myc, and LIN28 (2,3).
- Kim, J. et al. (2008) Cell 132, 1049-61.
- Takahashi, K. et al. (2007) Nat Protoc 2, 3081-9.
- Yu, J. et al. (2007) Science 318, 1917-20.
Application References
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For Research Use Only. Not For Use In Diagnostic Procedures.
