Invitrogen™ Nanog Monoclonal Antibody (eBioMLC-51), eFluor™ 660, eBioscience™, Invitrogen™

Catalog No.	Quantity
50-576-182	100 μg
50-576-180	25 μg

Catalog No. 50-576-182 Supplier Invitrogen™ Supplier No. 50576182

Description

Rat Monoclonal Antibody

The eBioMLC51 MAb 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, Cat. 00-5523, for intracellular staining of mouse Nanog.

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.

Specifications

• Antigen: Nanog
• Applications: Flow Cytometry, Immunohistochemistry, Western Blot, Immunocytochemistry
• Classification: Monoclonal
• Clone: eBioMLC-51
• Concentration: 0.2 mg/mL
• Conjugate: eFluor 660
• Formulation: PBS with 0.09% sodium azide; pH 7.2
• Gene: NANOG
• Gene Accession No.: Q80Z64
• Gene Alias: 2410002E02Rik; early embryo specific expression NK family; early embryo specific expression NK-type homeobox protein; Ecat4; ENK; ES cell-associated protein 4; FLJ12581; HGNC:20857; hNanog; Homeobox protein NANOG; homeobox transcription factor Nanog; homeobox transcription factor Nanog-delta 48; Nanog; Nanog homeobox
• Gene Symbols: NANOG
• Host Species: Rat
• Purification Method: Affinity chromatography
• Quantity: 100 μg
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 71950
• Target Species: Mouse
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG2a κ

Frequently Asked Questions (FAQs)

I am unable to analyze my cells stained with eFluor Organic Dyes today. What options do I have?

Our options will depend on the samples you are analyzing.

If cell viability is not critical, you can store your stained samples at 4 degrees C or on ice overnight in the dark and analyze the following day.

For samples stained with eFluor organic fluorochromes, we recommend that cells be suspended in 100 uL of Flow Cytometry Staining Buffer (Cat. No. 00-4222) and 100 uL of eBioscience IC Fixation Buffer (Cat. No. 00-8222); samples can be incubated for up to 3 days at 4 degrees C in the dark. Alternatively, the 1-step Fix/Lyse Solution (Cat. No. 00-5333) can be used. This is a great option when working with whole blood but also works for other cell types.

Can the eFluor Organic fluorochromes be used for intracellular staining?

Yes, the eFluor Organic fluorochromes can be used for intracellular staining. The eFluor organic fluorochromes maintain bright signal and require minimal changes in compensation when fixed with eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) and Permeabilization Buffer (Cat. No. 00-8333-56) or 1-step Fix/Lyse Solution (Cat. No. 00-5333-54, 00-5333-57) (as compared to live cells).

Is the eFluor 660 fluorochrome compatible with Anti-Cy5/Alexa Fluor 647 beads?

Yes, in-house studies have demonstrated that the eFluor 660 fluorochrome is recognized by Anti-Cy5/Alexa Fluor 647 beads. Side by side studies with Alexa Fluor 647 versus eFluor 660 conjugated antibodies have demonstrated comparable results.

What buffers are compatible with the eFluor Organic fluorochromes and eVolve QDots?

The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.

What is the difference between eFluor Organic Dyes and eVolve Qdots?

The eFluor Organic Dyes (eFluor 450, APC-eFluor 780, PerCP-eFluor 710, eFluor 710) are conventional fluorochromes. In contrast, the eVolve line of products are Quantum dots.

Are the eFluor Organic Dyes photo-labile?

As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.

Safety and Handling

WARNING: Cancer - www.P65Warnings.ca.gov