Rabbit Anti-Nanog antibody | Sunlong Medical

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NANOG_MOUSE; Embryonic stem cell specific homeobox protein (Nanog); ENK; FLJ12581; FLJ40451; Homeobox transcription factor Nanog; Nanog homeobox; Homeobox protein NANOG; ES cell-associated protein 4; Early embryo specific expression NK-type homeobox prote

Cat:

SL10414R

Species Reactivity：

Human,Mouse,Rat,

Immunogen：

KLH conjugated synthetic peptide derived from mouse Nanog:101-200/305

Format：

Liquid

Storage instructions：

Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Concentration：

1mg/ml

Clonality：

Polyclonal

Isotype：

IgG

Applications：

WB=1:500-2000ELISA=1:5000-10000IHC-P=1:100-500IHC-F=1:100-500ICC=1:100-500IF=1:100-500（Paraffin sections need to do antigen repair）not yet tested in other applications.optimal dilutions/concentrations should be determined by the end user.

Host：

Rabbit

Product Overview：

Sample: A431(Human) Cell Lysate at 30 ugPrimary: Anti-Nanog (SL10414R) at 1/300 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 34 kDObserved band size: 34 kDSample: NIH/3T3(Mouse) Cell Lysate at 30 ugPrimary: Anti-Nanog (SL10414R) at 1/300 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 34 kDObserved band size: 34 kDTissue/cell: mouse embryo tissue; 4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min; Incubation: Anti-Nanog Polyclonal Antibody, Unconjugated(SL10414R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) stainingTissue/cell: rat testis tissue; 4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min; Incubation: Anti-Nanog Polyclonal Antibody, Unconjugated(SL10414R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) stainingTissue/cell: rat testis tissue; 4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min; Incubation: Anti-Nanog Polyclonal Antibody, Unconjugated(SL10414R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining Paraformaldehyde-fixed, paraffin embedded (Mouse ovarian); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Nanog) Polyclonal Antibody, Unconjugated (SL10414R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Paraformaldehyde-fixed, paraffin embedded (Mouse ovary); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Nanog) Polyclonal Antibody, Unconjugated (SL10414R) at 1:400 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (SL0295G-CY3) for 90 minutes, and DAPI for nuclei staining.Blank control: Hela. Primary Antibody (green line): Rabbit Anti- Nanog antibody (SL10414R) Dilution: 3μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : Goat anti-rabbit IgG-PEDilution: 1μg /test. ProtocolThe cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with ice-cold methanol for 20 min at-20℃.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

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Unit:

Price: $228.00

Product PDFs

Datasheet: Down Datasheet

Other Information
Citations
Protein Attributes
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Nanog is a newly identified homeodomain-bearing transcriptional factor. Nanog expression is specific to early embryos and pluripotential stem cells including mouse and human embryonic stem (ES) and embryonic germ (EG) cells. It is a key molecule involved in the signaling pathway for maintaining the capacity for self-renewal and pluripotency, bypassing regulation by the STAT3 pathway. Nanog mRNA is present in pluripotent mouse and human cell lines, and absent from differentiated cells. Nanog-deficient ES cells lose pluripotency and differentiate into extraembryonic endoderm lineage. Thus it is one of the molecular markers suitable for recognizing the undifferentiated state of stem cells in the mouse and human.
NANOG is a new marker for testicular carcinoma in situ and germ cell tumors.
NANOG is a gene expressed in embryonic stem cells (ESCs) and is thought to be a key factor in maintaining pluripotency. NANOG thought to function in concert with other factors such as POU5F1 and SOX2 to establish ESC identity. These cells offer an important area of study because of their ability to maintain pluripotency. In other words, these cells have the ability to become virtually any cell of any of the three germ layers (endoderm, ectoderm, mesoderm).

Function:
Transcription 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. Acts as a transcriptional activator or repressor. Binds optimally to the DNA consensus sequence 5'-TAAT[GT][GT]-3' or 5'-[CG][GA][CG]C[GC]ATTAN[GC]-3'. When overexpressed, promotes cells to enter into S phase and proliferation.

Subunit:
Interacts with SMAD1 and SALL4.

Subcellular Location:
Nucleus.

Tissue Specificity:
Expressed in testicular carcinoma and derived germ cell tumors (at protein level). Expressed in fetal gonads, ovary and testis. Also expressed in ovary teratocarcinoma cell line and testicular embryonic carcinoma. Not expressed in many somatic organs and oocytes.

Similarity:
Belongs to the Nanog homeobox family.
Contains 1 homeobox DNA-binding domain.

SWISS:
Q80Z64

Gene ID:
71950

Database links:

Entrez Gene: 100293888 Human

Entrez Gene: 79923 Human

Entrez Gene: 71950 Mouse

Omim: 607937 Human

SwissProt: Q9H9S0 Human

Unigene: 635882 Human

Picture

Sample:
NIH/3T3(Mouse) Cell Lysate at 30 ug
Primary: Anti-Nanog (SL10414R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 34 kD
Observed band size: 34 kD

Tissue/cell: mouse embryo tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min;
Incubation: Anti-Nanog Polyclonal Antibody, Unconjugated(SL10414R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining

Tissue/cell: rat testis tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min;
Incubation: Anti-Nanog Polyclonal Antibody, Unconjugated(SL10414R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining

Paraformaldehyde-fixed, paraffin embedded (Mouse ovarian); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Nanog) Polyclonal Antibody, Unconjugated (SL10414R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Mouse ovary); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Nanog) Polyclonal Antibody, Unconjugated (SL10414R) at 1:400 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (SL0295G-CY3) for 90 minutes, and DAPI for nuclei staining.

Blank control: Hela.
Primary Antibody (green line): Rabbit Anti- Nanog antibody (SL10414R)
Dilution: 3μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-PE
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with ice-cold methanol for 20 min at-20℃.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

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