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    dkk1-antibody-epr4759-ab109416.pdf

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Immunology Immune Interventions Transplantation
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RecombinantRabMAb

Recombinant Anti-DKK1 antibody [EPR4759] (ab109416)

  • Datasheet
  • SDS
Reviews (1) Submit a question References (34)

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Western blot - Anti-DKK1 antibody [EPR4759] (ab109416)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DKK1 antibody [EPR4759] (ab109416)
  • Western blot - Anti-DKK1 antibody [EPR4759] (ab109416)
  • Western blot - Anti-DKK1 antibody [EPR4759] (ab109416)
  • Western blot - Anti-DKK1 antibody [EPR4759] (ab109416)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DKK1 antibody [EPR4759] (ab109416)
  • Anti-DKK1 antibody [EPR4759] (ab109416)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4759] to DKK1
  • Suitable for: WB, IHC-P
  • Reacts with: Human, Recombinant fragment

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Overview

  • Product name

    Anti-DKK1 antibody [EPR4759]
    See all DKK1 primary antibodies
  • Description

    Rabbit monoclonal [EPR4759] to DKK1
  • Host species

    Rabbit
  • Specificity

    Although our internal data shows that ab109416 is not suitable to detect DKK1 in rats, ab109416 has been referenced in PMID 29299122 and 25469240 to work with rat derived cells. Please contact technical@abcam.com for further information.
  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt or ICC/IF
  • Species reactivity

    Reacts with: Human, Recombinant fragment
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HEK-293, Caco-2, U-2 OS, MOLT4, A549, PC-3, NCI-H460, DU145 and SK-OV-3 whole cell lysates IHC-P: Human placenta and liver tissue
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR4759
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Immune Interventions
    • Transplantation
    • Cardiovascular
    • Blood
    • Other
    • Stem Cells
    • Signaling Pathways
    • Wnt
    • Secreted
    • Developmental Biology
    • Embryogenesis
    • Axis formation and symmetry

Associated products

  • Alternative Versions

    • Anti-DKK1 antibody [EPR4759] - Low endotoxin, Azide free (ab221779)
    • Anti-DKK1 antibody [EPR4759] - BSA and Azide free (ab271882)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant human DKK1 protein (ab155623)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab109416 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB (1)
1/1000 - 1/10000. Detects a band of approximately 38 kDa (predicted molecular weight: 29 kDa).
IHC-P
1/800. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

1/250.00000 - 1/500.00000

Notes
WB
1/1000 - 1/10000. Detects a band of approximately 38 kDa (predicted molecular weight: 29 kDa).
IHC-P
1/800. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

1/250.00000 - 1/500.00000

Application notes
Is unsuitable for Flow Cyt or ICC/IF.

Target

  • Function

    Antagonizes canonical Wnt signaling by inhibiting LRP5/6 interaction with Wnt and by forming a ternary complex with the transmembrane protein KREMEN that promotes internalization of LRP5/6. DKKs play an important role in vertebrate development, where they locally inhibit Wnt regulated processes such as antero-posterior axial patterning, limb development, somitogenesis and eye formation. In the adult, Dkks are implicated in bone formation and bone disease, cancer and Alzheimer disease.
  • Tissue specificity

    Placenta.
  • Sequence similarities

    Belongs to the dickkopf family.
  • Domain

    The C-terminal cysteine-rich domain mediates interaction with LRP5 and LRP6.
  • Cellular localization

    Secreted.
  • Target information above from: UniProt accession O94907 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 22943 Human
    • Omim: 605189 Human
    • SwissProt: O94907 Human
    • Unigene: 40499 Human
    • Alternative names

      • Dickkopf 1 antibody
      • Dickkopf 1 homolog antibody
      • Dickkopf 1 like antibody
      • Dickkopf homolog 1 antibody
      • Dickkopf like protein 1 antibody
      • Dickkopf related protein 1 antibody
      • Dickkopf WNT signaling pathway inhibitor 1 antibody
      • Dickkopf-1 antibody
      • Dickkopf-related protein 1 antibody
      • DKK 1 antibody
      • Dkk-1 antibody
      • Dkk1 antibody
      • DKK1_HUMAN antibody
      • hDkk 1 antibody
      • hDkk-1 antibody
      • SK antibody
      see all

    Images

    • Western blot - Anti-DKK1 antibody [EPR4759] (ab109416)
      Western blot - Anti-DKK1 antibody [EPR4759] (ab109416)
      All lanes : Anti-DKK1 antibody [EPR4759] (ab109416) at 1/2000 dilution

      Lane 1 : Caco-2 cell lysate
      Lane 2 : PC-3 cell lysate
      Lane 3 : U-2 OS cell lysate
      Lane 4 : MOLT4 cell lysate
      Lane 5 : HEK-293 cell lysate

      Performed under reducing conditions.

      Predicted band size: 29 kDa
      Observed band size: 30-45 kDa why is the actual band size different from the predicted?



      False colour image of Western blot: Anti-DKK1 antibody [EPR4759] staining at 1/2000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DKK1 antibody [EPR4759] (ab109416)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DKK1 antibody [EPR4759] (ab109416)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human liver tissue sections labeling DKK1 with Purified ab109416 at 1:800 dilution (1.04 µg/ml). Heat mediated antigen retrieval was performed using using ab93684 (Tris/EDTA buffer, pH 9.0)ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody.Negative control:PBS instead of the primary antibody.Hematoxylinwas used as a counterstain
    • Western blot - Anti-DKK1 antibody [EPR4759] (ab109416)
      Western blot - Anti-DKK1 antibody [EPR4759] (ab109416)
      All lanes : Anti-DKK1 antibody [EPR4759] (ab109416) at 1/10000 dilution

      Lane 1 : Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate
      Lane 2 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 29 kDa
      Observed band size: 29-40 kDa why is the actual band size different from the predicted?


      Exposure time: 180 seconds


      Blocking/Diluting buffer and concentration 5% NFDM/TBST.

      HEK-293 and 293T cell are weak expressed cell lines for DKK1 protein (PMID: 18502762,  PMID: 15378020).

    • Western blot - Anti-DKK1 antibody [EPR4759] (ab109416)
      Western blot - Anti-DKK1 antibody [EPR4759] (ab109416)
      Anti-DKK1 antibody [EPR4759] (ab109416) at 1/1000 dilution + His-Tagged human DKK1 (aa 32 to 266) recombinant protein, 100ng

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 29 kDa



      Blocking/Diluting buffer and concentration 5% NFDM /TBST

      Exposure time: 5.5 seconds

       

    • Western blot - Anti-DKK1 antibody [EPR4759] (ab109416)
      Western blot - Anti-DKK1 antibody [EPR4759] (ab109416)
      All lanes : Anti-DKK1 antibody [EPR4759] (ab109416) at 1/2000 dilution (purified)

      Lane 1 : A549 (Human lung carcinoma epithelial cell) whole cell lysates
      Lane 2 : PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysates
      Lane 3 : NCI-H460 (Human Large Cell Lung Cancer epithelial cell) whole cell lysates
      Lane 4 : DU145 (Human prostate carcinoma epithelial cell) whole cell lysates
      Lane 5 : SK-OV-3 (Human ovarian cancer epithelial cell) whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 29 kDa



      Blocking and diluting buffer: 5% NFDM/TBST.

      The smear bands detected by ab109416 are similar to those reported by the papers (PMID: 18546262, 28090290, 24008318), which should be due to the glycosylation of DKK1. 

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DKK1 antibody [EPR4759] (ab109416)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DKK1 antibody [EPR4759] (ab109416)

      Immunohistochemical analysis of paraffin-embedded Human placenta tissue using 1/100 unpurified ab109416.

    • Anti-DKK1 antibody [EPR4759] (ab109416)
      Anti-DKK1 antibody [EPR4759] (ab109416)

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (34)

    Publishing research using ab109416? Please let us know so that we can cite the reference in this datasheet.

    ab109416 has been referenced in 34 publications.

    • Shailesh H  et al. Protein arginine methyltransferase 5 (PRMT5) activates WNT/ß-catenin signalling in breast cancer cells via epigenetic silencing of DKK1 and DKK3. J Cell Mol Med 25:1583-1600 (2021). PubMed: 33462997
    • Qi L  et al. HomeoboxC6 promotes metastasis by orchestrating the DKK1/Wnt/ß-catenin axis in right-sided colon cancer. Cell Death Dis 12:337 (2021). PubMed: 33795652
    • Zhao Y  et al. Andrographis overcomes 5-fluorouracil-associated chemoresistance through inhibition of DKK1 in colorectal cancer. Carcinogenesis 42:814-825 (2021). PubMed: 33822896
    • Zhou G  et al. The regulation effect of WNT-RAS signaling in hypothalamic paraventricular nucleus on renal fibrosis. J Nephrol 33:289-297 (2020). PubMed: 31392659
    • Bak DH  et al. Protective effects of human umbilical cord blood-derived mesenchymal stem cells against dexamethasone-induced apoptotic cell death in hair follicles. Int J Mol Med 45:556-568 (2020). PubMed: 31894311
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Western blot abreview for Anti-DKK1 antibody [EPR4759]

    Below Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HUVEC)
    Loading amount
    20 µg
    Specification
    HUVEC
    Gel Running Conditions
    Reduced Denaturing (8% gel)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 16 2011

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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