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Nanog Monoclonal Antibody (eBioMLC-51), Alexa Fluor™ 488, eBioscience™
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FIGURE: 1 / 80
Nanog Antibody (53-5761-80) in Flow
F9 cells were stained intracellularly, using the Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523-00) and protocol, with 0.06 µg of Rat IgG2a kappa Isotype Control, Alexa Fluor 488 (Product # 53-4321-80) (blue histogram) or 0.06 µg of Nanog Monoclonal Antibody, Alexa Fluor 488 (purple histogram). Total cells were used for analysis.
Please note: We are reviewing Western blot images included in the antibody testing data in our catalog, including those provided by third parties. Unless expressly labeled or annotated as “raw-unedited”, Western blot images included in the antibody testing data in our catalog may have been edited, optimized or otherwise adjusted for presentation.
Product Details
53-5761-80
Applications
Tested Dilution
Publications
Product Specifications
Species Reactivity
Mouse
Published species
Mouse,
Rat
Host/Isotype
Rat
/ IgG2a, kappa
Recommended Isotype Control
Class
Monoclonal
Type
Antibody
Clone
eBioMLC-51
Conjugate
Alexa Fluor™ 488
Alexa Fluor™ 488
Alexa Fluor™ 488
Excitation/Emission Max
499/520 nm
View spectra
Form
Liquid
Concentration
0.5 mg/mL
Amount
25 µg
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.2
Contains
0.09% sodium azide
Storage conditions
4°C, store in dark, DO NOT FREEZE!
Shipping conditions
Ambient (domestic); Wet ice (international)
RRID
Product Specific Information
Description: The eBioMLC51 monoclonal antibody recognizes mouse Nanog. Nanog is a multidomain homeobox transcription factor that has been shown to maintain pluripotency of embryonic stem cells, independent of LIF/Stat3. Expression of Nanog in the mouse is specific to early embryos, the ICM of the blastocyst, embryonic stem (ES) cells, and embryonic germ (EG) cells. Nanog expression often overlaps, but is not identical to, that of Oct4. Nanog is downregulated upon cellular differentiation and loss of pluripotency, making it a suitable marker in determining the undifferentiated state of stem cells.
Nanog acts as a transcriptional activator and has two activation domains in the C-terminus, called CD2 and WR, and one activation domain in the N terminus. The CD2 domain is unique to Nanog, whereas the NK2 DNA binding domain is well conserved.
Immunoblotting using eBioMLC51 reveals a band at ~ 45 kDa in F9 (an embryonal carcinoma cell line) lysate, but not in lysate from the NIH3T3 cell line or mouse spleen.
Preliminary data using fluorochrome-conjugated eBioMLC51 suggests that it is essential to use the eBioscience Foxp3 Staining Buffer Set, Product # 00-5523-00, for intracellular staining of mouse Nanog.
Applications Reported: This eBioMLC-51 antibody has been reported for use in intracellular staining followed by flow cytometric analysis, and immunocytochemistry.
Applications Tested: This eBioMLC-51 antibody has been tested by intracellular staining followed by flow cytometric analysis of the F9 cell line using the Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523) and protocol. Please refer to BestProtocols®: Protocol B: One-step protocol for intracellular (nuclear) proteins. This antibody can be used at less than or equal to 0.06 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. This eBioMLC-51 antibody has been tested by immunocytochemistry on fixed and permeabilized F9 cells and can be used at less than or equal to 10 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Excitation: 488 nm; Emission: 519 nm; Laser: Blue Laser.
Filtration: 0.2 µm post-manufacturing filtered.
Target Information
NANOG (Nanog homeobox) is a divergent homeodomain protein that directs pluripotency and differentiation of undifferentiated embryonic stem cells. NANOG mRNA is present in pluripotent mouse and human cell lines, and absent from differentiated cells. Human NANOG protein shares 52% overall amino acid identity with the mouse protein, and 85% identity in the homeodomain. Human NANOG maps to gene locus 12p13.31, while the mouse NANOG maps to gene loci 6 F2. Murine embryonic NANOG expression is detected in the inner cell mass of the blastocyst. Research studies have shown that high levels of human NANOG expression in the undifferentiated N-Tera embryonal carcinoma cell line. Further, NANOG is a transcription regulator involved in inner cell mass and embryonic stem (ES) cells proliferation and self-renewal. The role of NANOG in embryonic development suggested that it might be useful in the creation of stem cells that might be useful in cell replacement therapies in the treatment of several degenerative diseases. Artificial stem cells, termed induced pluripotent stem (iPS) cells, can be created by expressing POU5F1 (also known as Oct-4), another germline-specific transcription factor, and the transcription factors Sox2, Klf4 and Lin28 along with c-Myc in mouse fibroblasts. Experiments have demonstrated that iPS cells could be generated using expression plasmids expressing NANOG, Sox2, KlfF4 and c-Myc, eliminating the need for virus introduction, thereby addressing a safety concern for potential use of iPS cells in regenerative medicine. When overexpressed, NANOG promotes cells to enter into S phase and proliferation. Diseases associated with dysfunction in the NANOG protein include tetracarcinoma and germ cell/embryonal cancer.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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Bioinformatics
Protein Aliases: early embryo specific expression NK family; early embryo specific expression NK-type homeobox protein; ES cell-associated protein 4; ES specific homeobox protein; FLJ12581; HGNC:20857; homeobox transcription factor Nanog; homeodomain protein; transcript variant 1a; transcript variant 1b; unnamed protein product
Gene Aliases: 2410002E02Rik; ecat4; ENK; Stm1
UniProt ID: (Mouse) Q80Z64
Entrez Gene ID: (Mouse) 71950
transcription regulatory region sequence-specific DNA binding
RNA polymerase II regulatory region sequence-specific DNA binding
RNA polymerase II core promoter proximal region sequence-specific DNA binding
RNA polymerase II transcription factor activity, sequence-specific DNA binding
transcriptional repressor activity, RNA polymerase II transcription regulatory region sequence-specific binding
transcriptional activator activity, RNA polymerase II transcription regulatory region sequence-specific binding
DNA binding
chromatin binding
transcription factor activity, sequence-specific DNA binding
protein binding
transcription factor binding
identical protein binding
sequence-specific DNA binding
sequence-specific double-stranded DNA binding
negative regulation of transcription from RNA polymerase II promoter
mesodermal cell fate commitment
endodermal cell fate specification
regulation of transcription, DNA-templated
regulation of transcription from RNA polymerase II promoter
positive regulation of cell proliferation
gonad development
embryonic pattern specification
negative regulation of cell fate commitment
gene expression
stem cell division
stem cell population maintenance
cell differentiation
negative regulation of BMP signaling pathway
response to retinoic acid
negative regulation of endodermal cell fate specification
cell dedifferentiation
negative regulation of cell differentiation
positive regulation of transcription, DNA-templated
positive regulation of mitotic cell cycle
positive regulation of transcription from RNA polymerase II promoter
stem cell differentiation
regulation of stem cell division
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