Anti-Nanog Antibody, clone 4C17 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1566 - recombinant, expressed in HEK 293 cells
Synonym: Homeobox protein NANOG; Homeobox transcription factor Nanog; hNanog
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human testis cancer (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1566, Anti-Nanog, clone 4C17 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Nuclear staining was observed in tumor cells and subsets of germ cells in adjacent seminiferous tubules of human testis cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human testis cancer (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1566, Anti-Nanog, clone 4C17 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Nuclear staining was observed in tumor cells and subsets of germ cells in adjacent seminiferous tubules of human testis cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human testis cancer (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1566, Anti-Nanog, clone 4C17 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Nuclear staining was observed in tumor cells and subsets of germ cells in adjacent seminiferous tubules of human testis cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human testis cancer (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1566, Anti-Nanog, clone 4C17 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Nuclear staining was observed in tumor cells and subsets of germ cells in adjacent seminiferous tubules of human testis cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human testis cancer (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1566, Anti-Nanog, clone 4C17 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Nuclear staining was observed in tumor cells and subsets of germ cells in adjacent seminiferous tubules of human testis cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human testis cancer (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1566, Anti-Nanog, clone 4C17 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Nuclear staining was observed in tumor cells and subsets of germ cells in adjacent seminiferous tubules of human testis cancer tissue sections.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of NTERA-2 cell line was performed using a 1:100 dilution of Cat. No. ZRB1566, Anti-Nanog, clone 4C17 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of NTERA-2 cell line was performed using a 1:100 dilution of Cat. No. ZRB1566, Anti-Nanog, clone 4C17 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of NTERA-2 cell line was performed using a 1:100 dilution of Cat. No. ZRB1566, Anti-Nanog, clone 4C17 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Western Blotting Enhanced Validation-Recombinant Antibody Technology NTERA-2 cell nuclear extract (Lane 1) and H9 cell lysate (Lane 2) were probed with Cat. No. ZRB1566, Anti-Nanog, clone 4C17 ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates Nanog (~42 kDa).
Affinity Binding Assay Enhanced Validation-Recombinant Antibody Technology 100 μg/mL of of non-specific GST protein (Run 1) or human recombinant Nanog protein (Run2) were analyzed by affinity binding assay using ZRB1566, Anti-Nanog, clone 4C17 ZooMAb® Rabbit Monoclonal. The binding of this antibody to recombinant Nanog protein displayed a KD value of 7.6 x 10-9 whereas its binding to non-specific GST protein displayed a KD of 3.9 x 10-5.
Flow Cytometry Enhanced Validation-Recombinant Antibody Technology Staining of one million NTERA-2 cells was performed using 1 μg of Cat. No. ZRB1566, Anti-Nanog, clone 4C17 ZooMAb® Rabbit Monoclonal (Yellow histogram) or the equivalent amount of a Rabbit IgG isotype control (Grey histogram), followed by a PE-conjugated Donkey Anti-Rabbit secondary antibody.
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | rabbit |
| clone | 4C17, monoclonal |
| recombinant monoclonal | |
| conjugate | unconjugated |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG |
| mol wt | calculated mol wt 34.62 kDa |
| observed mol wt ~42 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| flow cytometry: suitable | |
| immunocytochemistry: suitable | |
| immunohistochemistry: suitable | |
| western blot: suitable | |
| UniProt accession no. | Q9H9S0  |
| Application: | Anti-Nanog, clone 4C17 ZooMAb, Cat. No. ZRB1566, is a recombinant Rabbit monoclonal antibody that targets Nanog and is tested for use in Affinity Binding Assay, Flow Cytometry, Immunocytochemistry, Immunohistochemistry, and Western Blotting. |
| Application: | Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected Nanog in H9 cell lysate. Flow Cytometry Analysis: 1 μg from a representative lot detected Nanog in one million NTERA-2 cells. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected Nanog in NTERA-2 cells. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected Nanog in human testis cancer tissue sections. Affinity Binding Assay: A representative lot of this antibody bound human recombinant Nanog with a KD of 7.6 x 10-9 in an affinity binding assay. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb antibodies are reliably available and ready to ship when you need them. Learn more about ZooMAb here. |
| Immunogen: | His-tagged full-length recombinant human Nanog. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 30 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 4C17 is a ZooMAb rabbit recombinant monoclonal antibody that specifically detects human Nanog. |
| Storage and Stability: | Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws. |
| Target description: | Homeobox protein Nanog (UniProt: Q9H9S0; Homeobox transcription factor Nanog, hNanog) is encoded by the NANOG gene (Gene ID: 79923) in human. Nanog is a transcription factor that is involved in the self-renewal of embryonic stem cells (ES) and is a critical factor for the maintenance of the undifferentiated state of pluripotent cells. Along with OCT4 and Sox2 it is responsible for maintaining ES cells in an undifferentiated state. It is expressed in embryonic stem (ES) and carcinoma cells and its expression is observed in inner cell mass (ICM) of the blastocyst and gonocytes between 14 and 19 weeks of gestation. Its expression decreases with ES differentiation. Nanog can autorepress its own expression in differentiating ES cells by binding to its own promoter following interaction with ZNF281/ZFP281, leading to recruitment of the NuRD complex and subsequent repression of expression. Overexpression of Nanog in ES cells maintains specific differentiation, however, its overexpression in cancer stem cells is reported to inhibit apoptosis. Nanog together with OCT4 can activate SNAIL2 and promote metastasis and the EMT phenotype in cancer cells. Nanog can be phosphorylated on serine 52, 71, and 78 and this can regulate its stability. Suppression of MEK/ERKs signaling is shown to enhance Nanog transactivation to inhibit ES cell differentiation. This ZooMAb recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Gawlik-Rzemieniewska, N., and Bednarek, I (2016). Cancer Biol. Ther. 17(1); 1-10; Kim, S-H., et al. (2014). Stem Cell Res. 13(1); 1-11). |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |