Human Midkine Antibody

(4 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human Midkine in direct ELISAs and Western blots. In direct ELISAs, approximately 10% cross-reactivity with recombinant mouse Midkine is observed, and less than 1% cross-reactivity with recombinant human Pleiotrophin is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human Midkine
    Lys23-Asp143
    Accession # P21741
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    2 µg/mL
    See below
  • Simple Western
    100 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
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.
Data Examples
Detection of Human Midkine by Western Blot. Western blot shows lysates of
SH‑SY5Y human neuroblastoma cell line. PVDF membrane was probed with 2 µg/mL of Goat Anti-Human Midkine Antigen Affinity-purified Polyclonal Antibody (Catalog #
AF-258-PB) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Midkine at approximately 17 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunohistochemistry
Midkine in Human Prostate. Midkine was detected in immersion fixed paraffin-embedded sections of human prostate using Goat Anti-Human Midkine Antigen Affinity-purified Polyclonal Antibody (Catalog # AF‑258‑PB) at 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to stromal cell cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Immunohistochemistry
Midkine in Human Ovarian Array. Midkine was detected in immersion fixed paraffin-embedded sections of human ovarian array using Goat Anti-Human Midkine Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-258-PB) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human Midkine by Simple WesternTM. Simple Western lane view shows lysates of SH‑SY5Y human neuroblastoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Midkine at approximately 26 kDa (as indicated) using 100 µg/mL of Goat Anti-Human Midkine Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-258-PB) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the
12-230 kDa separation system.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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: Midkine

Midkine (MK) is a 15 kDa heparin-binding molecule originally cloned during a search for genes preferentially transcribed during retinoic acid (RA)-induced differentiation. Midkine belongs to a family of neurotrophic and developmentally-regulated heparin-binding molecules consisting of midkine, pleiotrophin (PTN/HBNF/OSF-1/HNGF-8) and the avian midkine homolog, RI-HB (for retinoic acid-inducible heparin-binding protein).

Midkine is a highly basic, nonglycosylated polypeptide that contains five intrachain disulfide bonds. The predicted molecular weight is approximately 13.3 kDa, based on a mature peptide length of 118 amino acid residues in the mouse and 121 amino acid residues in the human. Across species, MK shows 87% identity between the human and murine proteins. Between family members, human MK is approximately 50% identical to human PTN, with conservation of all 10 cysteines. Initial structure-function studies indicate that the C-terminal half of MK contains the principal heparin-binding site plus the molecule’s antigenicity and neurite-promoting sequences; while both the C- and N-termini are necessary for the molecule’s neurotrophic effects. Cells known to produce MK include endothelial cells, fetal astrocytes, renal proximal tubule epithelial cells and Wilms’ (kidney) tumor cells. MK has also been identified in the senile plaques of patients with Alzheimer’s disease. The pattern of expression of midkine during development strongly suggests a role for this factor both in epithelial-mesenchymal interactions and in development of the nervous system.

  • References:
    1. Bohlen, P. and I. Kovesdi (1991) Prog. Growth Factor Res. 3:143.
    2. Muramatsu, T. (1993) Int. J. Dev. Biol. 37:183.
  • Entrez Gene IDs:
    4192 (Human); 17242 (Mouse)
  • Alternate Names:
    Amphiregulin-associated protein; ARAP; MDK; MEK; Midgestation and kidney protein; midkine (neurite growth-promoting factor 2); Midkine; MK1; MKARAP; NEGF2; NEGF2FLJ27379; Neurite outgrowth-promoting factor 2; Neurite outgrowth-promoting protein
Related Research Areas
Citations:

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.

4 Citations: Showing 1 - 4
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Species
Applications
Sample Type
  1. Regulation of MDK expression in human cancer cells modulates sensitivities to various anticancer drugs: MDK overexpression confers to a multi-drug resistance.
    Authors: Kang HC, Kim IJ, Park HW, Jang SG, Ahn SA, Yoon SN, Chang HJ, Yoo BC, Park JG
    Cancer Lett., 2007;247(1):40-7.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  2. Role of heparin binding growth factors in nigrostriatal dopamine system development and Parkinson's disease.
    Authors: Marchionini DM, Lehrmann E, Chu Y, He B, Sortwell CE, Becker KG, Freed WJ, Kordower JH, Collier TJ
    Brain Res., 2007;1147(0):77-88.
    Species: Rat
    Sample Type: Tissue Homogenates
    Application: WB
  3. Neuritogenic activity of chondroitin/dermatan sulfate hybrid chains of embryonic pig brain and their mimicry from shark liver. Involvement of the pleiotrophin and hepatocyte growth factor signaling pathways.
    Authors: Li F, Shetty AK, Sugahara K
    J. Biol. Chem., 2007;282(5):2956-66.
    Species: Mouse
    Sample Type: Whole Cells
    Application: Neut
  4. The anti-HIV cytokine midkine binds the cell surface-expressed nucleolin as a low affinity receptor.
    Authors: Said EA, Krust B, Nisole S, Svab J, Briand JP, Hovanessian AG
    J. Biol. Chem., 2002;277(40):37492-502.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
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