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Human/Mouse Dkk-1 Antibody

Catalog #: AF1096
21 Citations 1 Review Datasheet / COA / SDS
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AF1096
AF1096-SP
Detection of Human Dkk‑1 by Western Blot.
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Product Details
Citations (21)
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Reviews (1)
Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Mouse Dkk-1 Antibody Summary

Species Reactivity
Human, Mouse
Specificity
Detects human and mouse Dkk-1 in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant human (rh) Dkk-2, and rhDkk-4 is observed.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
S. frugiperda insect ovarian cell line Sf 21-derived recombinant human Dkk-1
Thr32-His266
Accession # O94907
Formulation
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.
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Blockade of Receptor-ligand Interaction
In a functional ELISA, 0.1-0.4 μg/mL of this antibody will block 50% of the binding of 50 ng/mL of recombinant human biotinylated Dkk-1 to immobilized recombinant human LRP-6 Fc Chimera (Catalog # 1505-LR) coated at 1 µg/mL (100 µL/well). At 10 μg/mL, this antibody will block >90% of the binding.
 
Knockout Validated
Dkk‑1 is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in Dkk‑1 knockout HeLa cell line.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human Dkk-1 antibody by Western Blot. View Larger

Detection of Human Dkk‑1 by Western Blot. Western blot shows lysates of MCF-7 human breast cancer cell line, HepG2 human hepatocellular carcinoma cell line, A431 human epithelial carcinoma cell line, and A549 human lung carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse Dkk-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1096) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for Dkk-1 at approximately 28-40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Western Blot Detection of Mouse Dkk-1 antibody by Western Blot. View Larger

Detection of Mouse Dkk‑1 by Western Blot. Western blot shows lysates of Neuro-2A mouse neuroblastoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse Dkk-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1096) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for Dkk-1 at approximately 28-40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Knockout Validated Western Blot Shows Human Dkk-1 Antibody Specificity by Using Knockout Cell Line. View Larger

Western Blot Shows Human Dkk‑1 Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Dkk-1 knockout HeLa cell line (KO). PVDF membrane was probed with 0.2 µg/mL of Goat Anti-Human/Mouse Dkk-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1096) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Dkk-1 at approximately 35 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & 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.

Background: Dkk-1

Dickkopf related protein 1 (Dkk-1) is a member of the Dkk protein family that includes Dkk-1, -2, -3, and -4 (1). All four members are secreted proteins that are synthesized as precursor proteins with an N-terminal signal peptide and 2 conserved cysteine-rich domains, which are separated by a linker region. Dkk proteins have potential furin type proteolytic cleavage sites, and short forms of Dkk-2 and Dkk-4 containing only the second cysteine-rich domain can be generated by proteolytic processing (1). Dkk proteins have distinct patterns of expression in adult and embryonic tissues, suggesting that they may play diverse roles in these tissues. The Dkk proteins have distinct effects on Wnt signaling. Dkk-1 and Dkk-4 are Wnt antagonists. Dkk-3 has not been demonstrated to affect Wnt signaling, and Dkk-2 acts as an agonist or antagonist, depending on the cellular context. Wnt signaling regulates many important developmental processes including neural crest differentiation, brain development, kidney morphogenesis, and sex determination. In addition, Wnt signaling has also been implicated in tumor formation. Canonical Wnt signaling via the beta-catenin pathway is transduced by a receptor complex composed of the Frizzled proteins (Fz) and low-density lipoprotein (LDL) receptor-related proteins (LRP5/6) (2, 3). Unlike many soluble Wnt antagonists that function by binding extracellular Wnt ligands to prevent interaction of Wnt with the Fz-LRP5/6 receptor complex, Dkk-1 and Dkk-4 antagonize Wnt/ beta -catenin signaling by direct high-affinity binding to the Wnt coreceptor LRP5/6 and inhibiting interaction of LRP5/6 with the Wnt-Frizzled complex (4). Dkk-1 and Dkk-4 also bind the transmembrane proteins Kremen1 (Krm1) and Krm2 with high-affinity (5). Krm2 has been shown to form a ternary complex with Dkk-1 or -4 and LRP5/6 to trigger internalization of the complex and removal LRP6 from the cell surface. Thus, Dkk-1/4 and Kremens function synergistically to antagonize LRP5/6-mediated Wnt activity. Dkk-2 also binds to LRP5/6 and the Kremens, but Dkk-2 acts as antagonist of the Wnt signaling pathway only in the presence of Krm2 (5, 6). Dkk 2 binding to LRP5/6 in the absence of Krm2 activates rather than inhibits Wnt signaling (6).

References
  1. Krupnik, V.E. et al. (1999) Gene 238:301.
  2. Zorn, A.M (2001) Current Biology R592.
  3. Mao, J. et al. (2001) Mol. Cell 7:801.
  4. Nusse, R. et al. (2001) Nature 411:255.
  5. Mao, J. et al. (2002) Nature 417:664.
  6. Mao, B. and C. Niehrs (2003) Gene 302:179.
Long Name
Dickkopf-1
Entrez Gene IDs
22943 (Human); 13380 (Mouse); 102123735 (Cynomolgus Monkey)
Alternate Names
dickkopf (Xenopus laevis) homolog 1; dickkopf homolog 1 (Xenopus laevis); dickkopf related protein-1; Dickkopf-1; dickkopf-related protein 1; Dkk1; Dkk-1; hDkk-1; SKdickkopf-1 like

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Citations for Human/Mouse Dkk-1 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

21 Citations: Showing 1 - 10
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  1. The m6A(m)-independent role of FTO in regulating WNT signaling pathways
    Authors: Hyunjoon Kim, Soohyun Jang, Young-suk Lee
    Life Science Alliance
  2. Dickkopf-1 Acts as a Profibrotic Mediator in Progressive Chronic Kidney Disease
    Authors: YC Hsu, CC Chang, CC Hsieh, YT Huang, YH Shih, HC Chang, PJ Chang, CL Lin
    International Journal of Molecular Sciences, 2023-04-21;24(8):.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  3. CCN1 interacts with integrins to regulate intestinal stem cell proliferation and differentiation
    Authors: JH Won, JS Choi, JI Jun
    Nature Communications, 2022-06-03;13(1):3117.
    Species: Mouse
    Sample Types: Organoids
    Applications: Neutralizing
  4. Dual Pharmacological Targeting of HDACs and PDE5 Inhibits Liver Disease Progression in a Mouse Model of Biliary Inflammation and Fibrosis
    Authors: A Claveria-C, L Colyn, I Uriarte, MU Latasa, M Arechederr, JM Herranz, L Alvarez, JM Urman, ML Martinez-C, JM Banales, B Sangro, K Rombouts, J Oyarzabal, JJG Marin, C Berasain, MA Avila, MG Fernandez-
    Cancers, 2020-12-13;12(12):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  5. CSN5/JAB1 suppresses the WNT inhibitor DKK1 in colorectal cancer cells
    Authors: S Jumpertz, T Hennes, Y Asare, AK Schütz, J Bernhagen
    Cell. Signal, 2017-02-14;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  6. Bulk cell density and Wnt/TGFbeta signalling regulate mesendodermal patterning of human pluripotent stem cells
    Nat Commun, 2016-12-09;7(0):13602.
    Species: Human
    Sample Types: Cell Culture Supernates
    Applications: Western Blot
  7. Parathyroid hormone-related protein inhibits DKK1 expression through c-Jun-mediated inhibition of beta-catenin activation of the DKK1 promoter in prostate cancer.
    Authors: Zhang H, Yu C, Dai J, Keller J, Hua A, Sottnik J, Shelley G, Hall C, Park S, Yao Z, Zhang J, McCauley L, Keller E
    Oncogene, 2013-06-10;33(19):2464-77.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  8. Wnt-beta-catenin-Tcf-4 signalling-modulated invasiveness is dependent on osteopontin expression in breast cancer.
    Authors: Ravindranath A, Yuen HF, Chan KK, Grills C, Fennell DA, Lappin TR, El-Tanani M
    Br. J. Cancer, 2011-07-19;105(4):542-51.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  9. Paracrine and autocrine signals induce and maintain mesenchymal and stem cell states in the breast.
    Authors: Scheel C, Eaton E, Li S, Chaffer C, Reinhardt F, Kah K, Bell G, Guo W, Rubin J, Richardson A, Weinberg R
    Cell, 2011-06-10;145(6):926-40.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  10. Expression pattern of REIC/Dkk-3 in various cell types and the implications of the soluble form in prostatic acinar development.
    Authors: 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
    Int. J. Oncol., 2010-12-01;37(6):1495-501.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  11. The fibroblast-derived paracrine factor neuregulin-1 has a novel role in regulating the constitutive color and melanocyte function in human skin.
    Authors: Choi W, Wolber R, Gerwat W
    J. Cell. Sci., 2010-08-24;123(0):3102-11.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  12. A potential role for Dkk-1 in the pathogenesis of osteosarcoma predicts novel diagnostic and treatment strategies.
    Authors: Lee N, Smolarz AJ, Olson S, David O, Reiser J, Kutner R, Daw NC, Prockop DJ, Horwitz EM, Gregory CA
    Br. J. Cancer, 2007-11-06;97(11):1552-9.
    Species: Human
    Sample Types: Cell Culture Supernates, Cell Lysates, Whole Tissue
    Applications: IHC-P, Immunodepletion, Western Blot
  13. Derivation of sarcomas from mesenchymal stem cells via inactivation of the Wnt pathway.
    Authors: Matushansky I, Hernando E, Socci ND, Mills JE, Matos TA, Edgar MA, Singer S, Maki RG, Cordon-Cardo C
    J. Clin. Invest., 2007-11-01;117(11):3248-57.
    Species: Human
    Sample Types: Cell Lysates, Whole Tissue
    Applications: IHC-P, Western Blot
  14. Wnt signaling antagonists are potential prognostic biomarkers for the progression of radiographic hip osteoarthritis in elderly Caucasian women.
    Authors: Lane NE, Nevitt MC, Lui LY, de Leon P, Corr M
    Arthritis Rheum., 2007-10-01;56(10):3319-25.
    Species: Human
    Sample Types: Serum
    Applications: ELISA Development
  15. Low-density lipoprotein receptor-related protein 5 (LRP5) mediates the prostate cancer-induced formation of new bone.
    Authors: Li ZG, Yang J, Vazquez ES, Rose D, Vakar-Lopez F, Mathew P, Lopez A, Logothetis CJ, Lin SH, Navone NM
    Oncogene, 2007-08-13;27(5):596-603.
    Species: Human, Mouse
    Sample Types: Cell Lysates, Whole Tissue
    Applications: IHC, Western Blot
  16. Thalidomide induces limb deformities by perturbing the Bmp/Dkk1/Wnt signaling pathway.
    Authors: Knobloch J, Shaughnessy JD, Ruther U
    FASEB J., 2007-02-05;21(7):1410-21.
    Species: Chicken
    Sample Types: Whole Cells
    Applications: Neutralization
  17. Differential gene expression in cultured osteoblasts and bone marrow stromal cells from patients with Paget's disease of bone.
    Authors: Naot D, Bava U, Matthews B, Callon KE, Gamble GD, Black M, Song S, Pitto RP, Cundy T, Cornish J, Reid IR
    J. Bone Miner. Res., 2007-02-01;22(2):298-309.
    Species: Human
    Sample Types: Serum
    Applications: ELISA Development
  18. Genomics identifies medulloblastoma subgroups that are enriched for specific genetic alterations.
    Authors: Thompson MC, Fuller C, Hogg TL, Dalton J, Finkelstein D, Lau CC, Chintagumpala M, Adesina A, Ashley DM, Kellie SJ, Taylor MD, Curran T, Gajjar A, Gilbertson RJ
    J. Clin. Oncol., 2006-03-27;24(12):1924-31.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  19. Inducible Stem-Cell-Derived Embryos Capture Mouse Morphogenetic Events In Vitro
    Authors: Gianluca Amadei, Kasey Y.C. Lau, Joachim De Jonghe, Carlos W. Gantner, Berna Sozen, Christopher Chan et al.
    Developmental Cell
  20. p38 MAPK regulates the Wnt inhibitor Dickkopf-1 in osteotropic prostate cancer cells
    Authors: A J Browne, A Göbel, S Thiele, L C Hofbauer, M Rauner, T D Rachner
    Cell Death & Disease
  21. CCL3 in the bone marrow microenvironment causes bone loss and bone marrow adiposity in aged mice
    Authors: Degang Yu, Shuhong Zhang, Chao Ma, Sen Huang, Long Xu, Jun Liang et al.
    JCI Insight

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Human Dkk-1 Antibody
By Anonymous on 12/04/2016
Application: WB Sample Tested: Cell Lysates Species: Human

Silencing of DKK-1 in MCF-7 cells. Buffer: 5% BSA. Dilution: 1/1,000.

Western Blot Human Dkk-1 Antibody AF1096