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Nanog antibody (pAb)

Host / Isotype

Rabbit / IgG

Reactivity

Human, Mouse

Applications

ChIP, ChIP-Seq, ICC, IF, WB

Cat No : 61419,61420 61419

Synonyms

Nanog, Nanog homeobox, Homeobox transcription factor Nanog, homeobox protein NANOG, hNanog, homeobox transcription factor Nanog-delta 48, antibody, polyclonal, pAb, wb, western blotting, sample, immunofluorescence, immunocytochemistry, IF, IC

验证数据展示

  • Nanog antibody (pAb) tested by ChIP-Seq. ChIP was performed using the ChIP-IT High Sensitivity Kit (Cat. No. 53040) with 30 ug of chromatin from undifferentiated hESC cells and 3 ul of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 9 million sequence tags were mapped to identify Nanog binding sites. The image shows binding across a region of chromosome 12. You can view the complete data set in the UCSC Genome Browser, starting at this specific location, here.

    Nanog antibody (pAb) tested by ChIP-Seq. ChIP was performed using the ChIP-IT High Sensitivity Kit (Cat. No. 53040) with 30 ug of chromatin from undifferentiated hESC cells and 3 ul of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 9 million sequence tags were mapped to identify Nanog binding sites. The image shows binding across a region of chromosome 12. You can view the complete data set in the UCSC Genome Browser, starting at this specific location, here.

  • Nanog antibody (pAb) tested by Immunofluorescence Mouse embryonic stem cells (mESCs) grown on mouse embryonic fibroblast feeder cells (MEFs) were fixed with 4% paraformaldehyde for 10 minutes at room temperature. Cells were then permeabilized and blocked by incubating with Blocking Solution containing 5% serum/0.1% Triton X-100 in D-PBS for 2 hours at room temperature. Cells were then incubated with Nanog antibody (Catalog No. 61419, red) at 1:200 dilution overnight at 4°C, washed with D-PBS, and incubated for 2 hours at room temperature with goat anti-mouse Cy3 secondary antibody at 1:250 dilution. Cells were visualized using a Zeiss fluorescent microscope at 20X magnification. Images show that Nanog antibody specifically stains mESC colonies and does not stain MEFs. Absence of Nanog staining in a subset of cells within the colonies suggests differentiation. Scale bars, 100 μm.

    Nanog antibody (pAb) tested by Immunofluorescence Mouse embryonic stem cells (mESCs) grown on mouse embryonic fibroblast feeder cells (MEFs) were fixed with 4% paraformaldehyde for 10 minutes at room temperature. Cells were then permeabilized and blocked by incubating with Blocking Solution containing 5% serum/0.1% Triton X-100 in D-PBS for 2 hours at room temperature. Cells were then incubated with Nanog antibody (Catalog No. 61419, red) at 1:200 dilution overnight at 4°C, washed with D-PBS, and incubated for 2 hours at room temperature with goat anti-mouse Cy3 secondary antibody at 1:250 dilution. Cells were visualized using a Zeiss fluorescent microscope at 20X magnification. Images show that Nanog antibody specifically stains mESC colonies and does not stain MEFs. Absence of Nanog staining in a subset of cells within the colonies suggests differentiation. Scale bars, 100 μm.

  • Nanog (pAb) tested by Western blot. 20 ug of mouse Embryonic Stem Cell (ESC) extract was run on SDS-PAGE and probed with antibody at 2 ug/ml.

    Nanog (pAb) tested by Western blot. 20 ug of mouse Embryonic Stem Cell (ESC) extract was run on SDS-PAGE and probed with antibody at 2 ug/ml.

产品信息

Tested Applications ChIP, ChIP-Seq, ICC, IF, WB

Applications Validated by Active Motif: ChIP: 5 ul per ChIP ChIP-Seq: 5 ul each ICC/IF: 1:200 dilution WB*: 0.5 - 2 ug/ml ChIP-Seq validation was performed by Active Motif's Epigenetics Services; the complete data set is available in the UCSC Genome Browser by clicking here. *Note: Many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt / Sonication Protocol when preparing nuclear extracts for Western blot. Individual optimization may be required.
Tested Reactivity Human, Mouse
Host / Isotype Rabbit / IgG
Class Polyclonal
Type Antibody
Immunogen This antibody was raised against a peptide within the N-terminal region of mouse Nanog.
Full Name Nanog antibody (pAb)
Synonyms Nanog, Nanog homeobox, Homeobox transcription factor Nanog, homeobox protein NANOG, hNanog, homeobox transcription factor Nanog-delta 48, antibody, polyclonal, pAb, wb, western blotting, sample, immunofluorescence, immunocytochemistry, IF, IC
Molecular weight 45 kDa
GenBank accession numberNP_082292
RRIDAB_2750953
Purification Method Affinity Purified
Buffer Purified IgG in PBS with 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.
Storage Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

背景介绍

Nanog (Nanog homeobox) is a transcriptional regulator involved in inner cell mass and embryonic stem (ES) cells proliferation and self-renewal. Imposes pluripotency on ES cells and prevents their differentiation towards extraembryonic endoderm and trophectoderm lineages. Blocks bone morphogenetic protein-induced mesoderm differentiation of ES cells by physically interacting with SMAD1 and interfering with the recruitment of coactivators to the active SMAD transcriptional complexes (By similarity). Acts as a transcriptional activator or repressor (By similarity). Binds optimally to the DNA consensus sequence 5'-TAAT[GT][GT]-3' or 5'-[CG][GA][CG]C[GC]ATTAN[GC]-3' (By similarity). When overexpressed, promotes cells to enter into S phase and proliferation (By similarity).