Anti-DKK3 Antibody, clone 3H17 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1074 - recombinant, expressed in HEK 293 cells
Synonym: Dickkopf-3; Dickkopf-related protein 3; Dkk-3; hDkk-3
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human cerebral cortex (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 dilution of Cat. No. ZRB1074, Anti-DKK3, clone 3H17 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/punctate cytoplasmic/membranous staining was observed in neurons and neuropil of human cerebral cortex tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human cerebral cortex (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 dilution of Cat. No. ZRB1074, Anti-DKK3, clone 3H17 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/punctate cytoplasmic/membranous staining was observed in neurons and neuropil of human cerebral cortex tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human cerebral cortex (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 dilution of Cat. No. ZRB1074, Anti-DKK3, clone 3H17 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/punctate cytoplasmic/membranous staining was observed in neurons and neuropil of human cerebral cortex tissue sections.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of MCF-7 cell line was performed using a 1:1,000 dilution of Cat. No. ZRB1074, Anti-DKK3, clone 3H17 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488. Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the punctate cytoplasmic signal.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of MCF-7 cell line was performed using a 1:1,000 dilution of Cat. No. ZRB1074, Anti-DKK3, clone 3H17 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488. Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the punctate cytoplasmic signal.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of MCF-7 cell line was performed using a 1:1,000 dilution of Cat. No. ZRB1074, Anti-DKK3, clone 3H17 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488. Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the punctate cytoplasmic signal.
Western Blotting Enhanced Validation-Recombinant Antibody Technology Human Brain tissue lysate was probed with Cat. No. ZRB1074, Anti-DKK3, clone 3H17 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 DKK3 (~58 kDa).
Affinity Binding Assay Enhanced Validation-Recombinant Antibody Technology 100 μg/mL of non-specific GST protein (Run 1) or recombinant Dickkopf-3 (DKK3) protein (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1074, Anti-DKK3, clone 3H17 ZooMAb® Rabbit Monoclonal. The binding of this antibody to recombinant DKK3 displayed a KD value of 5.7 x 10-9 whereas its binding to non-specific GST protein displayed a KD of 5.3 x 10-4.
Flow Cytometry Enhanced Validation-Recombinant Antibody Technology Staining of one million MCF-7 cells was performed using 0.1 μg of Cat. No. ZRB1074, Anti-DKK3, clone 3H17 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 IgG 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 | 3H17, 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 38.39 kDa |
| observed mol wt ~58 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 |
| immunocytochemistry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | Q9UBP4  |
| Application: | Affinity Binding Assay: A representative lot of this antibody bound recombinant DKK3 protein with a KD of 5.7 x 10-9 in an affinity binding assay. Immunocytochemistry Analysis: A 1:1,000 dilution from a representative lot detected DKK3 in MCF-7 cells. Flow Cytometry Analysis: 0.1 μg from a representative lot detected DKK3 in one million MCF-7 cells. Immunohistochemistry (Paraffin) Analysis: A 1:1,000 dilution from a representative lot detected DKK3 in human cerebral cortex tissue sections. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Application: | Anti-DKK3, clone 3H17 ZooMAb, Cat. No. ZRB1074, is a recombinant Rabbit monoclonal antibody that targets DKK3 and is tested for use in Affinity Binding Assay, Flow Cytometry, Immunocytochemistry, Immunohistochemistry (Paraffin), and Western Blotting. |
| 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 recombinant fragment corresponding to 300 amino acids (almost full length mature protein) from human Dickkopf-related protein 3 (DKK3). |
| 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: | 300 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 3H17 is a ZooMAb rabbit recombinant monoclonal antibody that specifically detects human Dickkopf-related protein 3 (DKK3). |
| 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: | Dickkopf-related protein 3 (UniProt: Q9UBP4; also known as Dickkopf-3, Dkk-3, hDkk-3) is encoded by the DKK3 (also known as REIC, UNQ258/PRO295) gene (Gene ID: 27122) in human. DKK is a family of secreted glycoproteins that display Wnt inhibitory activity. They act as high affinity ligands for the transmembrane proteins Kremen1 and 2, which are known to modulate Wnt signaling. They play a role in vertebrate development via locally inhibiting Wnt-regulated processes such as antero posterior axial patterning, limb development, somitogenesis and eye formation. DKK3 is synthesized with a signal peptide (aa 1-21), which is subsequently cleaved off in the mature form. High expression of DKK3 is reported in heart, brain, and spinal cord. DKK3 is shown to antagonize canonical Wnt signaling by inhibiting LRP5/6 interaction with Wnt and by forming a ternary complex with Kremen protein, which is known to promote internalization of LRP5/6. The C-terminal cysteine-rich domain of DKK3 mediates its interaction with LRP5 and LRP6. DKK3 is shown to induce a redistribution of β-catenin to the cell membrane where β-catenin promotes cell-cell adhesion by binding to the cytoplasmic domain of E-cadherin to form adheren junction complex. This binding also prevents nuclear translocation of β-catenin that reduces Wnt-mediated LEF/TCF transactivation. DKK3 is proposed to act as a tumor suppressor protein and its over-expression is shown to suppress tumor growth. This ZooMAb recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Hoang, BH., et al. (2004). Cancer Res. 64(8); 2734-2339; Niehrs. C. (2006). Oncogene. 25(57); 7469-7481). |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |