Human Midkine Antibody Summary
Accession # P21741
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.
Midkine in HepG2 human hepatocellular carcinoma cell line. Midkine was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line (left panel; positive staining) and HL-60 human acute promyelocytic leukemia cell line (right panel; negative staining) using Mouse Anti-Human Midkine Monoclonal Antibody (Catalog # MAB2582) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Midkine in Human Seminoma (testicle tumor). Midkine was detected in immersion fixed paraffin-embedded sections of human seminoma (testicle tumor) using Mouse Anti-Human Midkine Monoclonal Antibody (Catalog # MAB2582) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in cancer cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Detection of Midkine by Immunoprecipitation. Immunoprecipitation was performed on concentrated culture media from HAP1 human near-haploid cell using 2.0 μg of Mouse Anti-Human Midkine Monoclonal Antibody (Catalog # MAB25823) pre-coupled to protein G or protein A beads. Immunoprecipitated Midkine was detected with Goat Anti-Human Midkine Polyclonal Antibody (AF-258-PB) at 1.0 μg/ml. 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).
Preparation and Storage
- 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.
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.
- Bohlen, P. and I. Kovesdi (1991) Prog. Growth Factor Res. 3:143.
- Muramatsu, T. (1993) Int. J. Dev. Biol. 37:183.
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