Human Midkine Antibody

Catalog # Availability Size / Price Qty
AF-258-PB
AF-258-SP
Detection of Human Midkine by Western Blot.
6 Images
Product Details
Citations (6)
FAQs
Supplemental Products
Reviews (1)

Human Midkine Antibody Summary

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 either lyophilized or 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
Immunoprecipitation
1 µg/mL
 Conditioned cell culture medium from HAP1 human near-haploid 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 Midkine antibody by Western Blot. View Larger

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 (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 antibody in Human Prostate by Immunohistochemistry (IHC-P). View Larger

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 (CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown 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 antibody in Human Ovarian Array by Immunohistochemistry (IHC-P). View Larger

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; CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Simple Western Detection of Human Midkine antibody by Simple Western<sup>TM</sup>. View Larger

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 (HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Knockout Validated View Larger

Western Blot Shows Human Midkine Specificity Using Knockout Cell Line. Western blot shows lysates of HAP1 human near-haploid cell line untreated (-) or treated (+) with 3.0 ug/ml Brefeldin A for 18 hr and Midkine knockout HAP1 cell line (KO). Nitrocellulose 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. A specific band was detected for Midkine at approximately 11 kDa (as indicated) in the parental HAP1 cell line, but is not detectable in knockout HAP1 cell line. The Ponceau stained transfer of the blot is shown. This experiment was conducted under reducing conditions. Image, protocol, and testing courtesy of YCharOS Inc. See ycharos.com for additional details.

Immunoprecipitation View Larger

Detection of Midkine by Immunoprecipitation. Immunoprecipitation was performed on concentrated culture media of HAP1 human near-haploid cell line using 2.0 μg of Mouse Anti-Human Midkine Monoclonal Antibody (Catalog # AF-258-PB) pre-coupled to protein G or protein A beads. Immunoprecipitated Midkine was detected with Rabbit Anti-Human Midkine Monoclonal Antibody (NBP2-66948 ) at 1/500. The Ponceau stained transfers of each blot are shown. SM=10% starting material; UB=10% unbound fraction; IP=immunoprecipitated. Image, protocol, and testing courtesy of YCharOS Inc. (ycharos.com).

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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

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Citations for Human Midkine 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.

6 Citations: Showing 1 - 6
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  1. Midkine Is a Potential Therapeutic Target of Tumorigenesis, Angiogenesis, and Metastasis in Non-Small Cell Lung Cancer
    Authors: DH Shin, JY Jo, SH Kim, M Choi, C Han, BK Choi, SS Kim
    Cancers (Basel), 2020;12(9):.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  2. Midkine Is a Potential Therapeutic Target of Tumorigenesis, Angiogenesis, and Metastasis in Non-Small Cell Lung Cancer
    Authors: DH Shin, JY Jo, SH Kim, M Choi, C Han, BK Choi, SS Kim
    Cancers (Basel), 2020;12(9):.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  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 Types: Whole Cells
    Applications: Neutralization
  4. 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 Types: Cell Lysates
    Applications: Western Blot
  5. 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 Types: Tissue Homogenates
    Applications: Western Blot
  6. 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 Types: Whole Cells
    Applications: ICC

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Human Midkine Antibody
By Anonymous on 06/19/2019
Application: ELISA Sample Tested: Serum and Plasma Species: Human

AF258 was used with MAB2581 as a matched pair in an Midkine LBA.