Dkk-3 Antibody [Unconjugated]

Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Dkk‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1118) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for Dkk‑3 at approximately 38 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Dkk‑3 was detected in immersion fixed paraffin-embedded sections of Human Brain Hippocampus using Goat Anti-Human Dkk‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1118) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in neurons. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

• Reactivity: Hu
• Applications: WB, IHC
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Dkk-3 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human Dkk-3
  Ala22-Ile350
  Accession # Q9UBP4
• Specificity: Detects human Dkk-3 in direct ELISAs and Western blots. In direct ELISAs, approximately 35% cross-reactivity with recombinant mouse Dkk-3 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: DKK3
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 3-15 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Dkk-3 Antibody [Unconjugated]

• dickkopf (Xenopus laevis) homolog 3
• dickkopf 3
• dickkopf homolog 3 (Xenopus laevis)
• Dickkopf-3
• dickkopf-related protein 3
• Dkk3
• Dkk-3
• hDkk-3
• regulated in glioma
• REIC
• REICdkk-3
• RIG
• RIG-like 5-6
• RIG-like 7-1

Background

Dkk-3, also known as REIC (Reduced Expansion in Immortalized Cells), is one of four numbered members of the Dickkopf family of Wnt antagonists (1). Dkk-3 is a secreted monomer expressed in many normal human tissues, most strongly in heart, brain and spinal cord (1, 2), and during early embryonic development in the mouse (3). N-glycosylation at up to four sites preceding or between two conserved cysteine-rich motifs results in expression of a 38 kDa glycoprotein (1, 4). The cysteine-rich motifs contain 10 cysteines each, with prokineticin and colipase families containing sequences similar to those of the second motif (1, 5). Human Dkk-3 shows 82%, 88%, 85%, and 53% amino acid (aa) identity with mouse, bovine, canine, and chick Dkk-3, respectively, and 37-45% aa identity with other human Dkk family members. Several lines of evidence implicate Dkk-3 as a negative growth regulator. Dkk-3 is downregulated in many tumors as compared to normal cells, sometimes by loss of heterozygosity (4, 6). Downregulation by CpG hypermethylation in acute lymphoblastic leukemia is correlated with faster progression and shorter survival (7). Release of cultured cells from serum starvation results in downregulation of Dkk-3 in late G1 phase of the cell cycle (6). Over-expression of
Dkk‑3 results in tumor cell-line-specific growth inhibition, induction of apoptosis, and decreased tumorigenicity in nude mice (2, 4, 6). The prototype Dickkopf member, Dkk-1, antagonizes Wnt family signaling by binding to Wnt receptors LRP5 and LRP6 (low-density lipoprotein receptor-related proteins) and promoting their internalization (1, 9, 10). Results are less straightforward for Dkk-3, where some studies show binding to LRP5/6 while others do not. These effects appear to be dependent on the cells and conditions used (1, 6-10).

1. Krupnik, V.E. et al. (1999) Gene 238:301.
2. Tsuji, T. et al. (2000) Biochem. Biophys. Res. Comm. 268:20.
3. Kemp, C. et al. (2005) Dev. Dyn. 233:1064.
4. Hsieh, S.-Y. et al. (2004) Oncogene 23:9183.
5. Bullock, C.M. et al. (2004) Mol. Pharmacol. 65:582.
6. Tsuji, T. et al. (2001) Biochem. Biophys. Res. Comm. 289:257.
7. Roman-Gomez, J. et al. (2004) Br. J. Cancer 91:707.
8. Hoang, B.H. et al. (2004) Cancer Res. 64:2734.
9. Caricasole, A. et al. (2003) J. Biol. Chem. 278:37024.
10. Mao, B. et al. (2001) Nature 411:321.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Dkk-3 Antibody (AF1118)(10)

Publications using AF1118

• QTT Nguyen, HS Park, TJ Lee, KM Choi, JY Park, D Kim, JH Kim, J Park, EJ Lee DKK3, Downregulated in Invasive Epithelial Ovarian Cancer, Is Associated with Chemoresistance and Enhanced Paclitaxel Susceptibility via Inhibition of the beta-Catenin-P-Glycoprotein Signaling Pathway Cancers, 2022;14(4):. 2022 [PMID: 35205672] (IHC, Human)
• N Chiyonobu, S Shimada, Y Akiyama, K Mogushi, M Itoh, K Akahoshi, S Matsumura, K Ogawa, H Ono, Y Mitsunori, D Ban, A Kudo, S Arii, T Suganami, S Yamaoka, Y Ogawa, M Tanabe, S Tanaka Fatty acid binding protein 4 (FABP4) overexpression in intratumoral hepatic stellate cells within hepatocellular carcinoma with metabolic risk factors Am. J. Pathol., 2018;0(0):. 2018 [PMID: 29454748] (IHC-P, Human)
• Loss of Dickkopf 3 Promotes the Tumorigenesis of Basal Breast Cancer PLoS ONE, 2016;11(7):e0160077. 2016 [PMID: 27467270] (Western Blot, Human)
• Riordan D, Varma S, West R, Brown P Automated Analysis and Classification of Histological Tissue Features by Multi-Dimensional Microscopic Molecular Profiling. PLoS ONE, 2015;10(7):e0128975. 2015 [PMID: 26176839] (IHC-P, Human)
• Xiang T, Li L, Yin X, Zhong L, Peng W, Qiu Z, Ren G, Tao Q Epigenetic silencing of the WNT antagonist Dickkopf 3 disrupts normal Wnt/beta-catenin signalling and apoptosis regulation in breast cancer cells. J Cell Mol Med, 2013;17(10):1236-46. 2013 [PMID: 23890219] (Western Blot, Human)
• Wang W, Zhu W, Xu XY The clinicopathological significance of REIC expression in colorectal carcinomas. Histol. Histopathol., 2012;27(6):735-43. 2012 [PMID: 22473694] (Western Blot, Human)
• Zhang K, Watanabe M, Kashiwakura Y, Li SA, Edamura K, Huang P, Yamaguchi K, Nasu Y, Kobayashi Y, Sakaguchi M, Ochiai K, Yamada H, Takei K, Ueki H, Huh NH, Li M, Kaku H, Na Y, Kumon H Expression pattern of REIC/Dkk-3 in various cell types and the implications of the soluble form in prostatic acinar development. Int. J. Oncol., 2010;37(6):1495-501. 2010 [PMID: 21042718] (ICC, Human)
• Choi W, Wolber R, Gerwat W The fibroblast-derived paracrine factor neuregulin-1 has a novel role in regulating the constitutive color and melanocyte function in human skin. J. Cell. Sci., 2010;123(0):3102-11. 2010 [PMID: 20736300] (Western Blot, Human)
• Choi W, Miyamura Y, Wolber R, Smuda C, Reinhold W, Liu H, Kolbe L, Hearing VJ Regulation of human skin pigmentation in situ by repetitive UV exposure: molecular characterization of responses to UVA and/or UVB. J. Invest. Dermatol., 2010;130(6):1685-96. 2010 [PMID: 20147966] (IHC-P, Human)
• Untergasser G, Steurer M, Zimmermann M, Hermann M, Kern J, Amberger A, Gastl G, Gunsilius E The Dickkopf-homolog 3 is expressed in tumor endothelial cells and supports capillary formation. Int. J. Cancer, 2008;122(7):1539-47. 2008 [PMID: 18033687] (ICC, Human)

Bioinformatics

• Gene Symbol: DKK3
• Uniprot:
  • Q9UBP4()