Midkine Antibody (1011522) [Unconjugated]

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 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 (Catalog # VC001). 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 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.

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).

• Reactivity: Hu
• Applications: IHC, IP, ICC/IF
• Clone: 1011522
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

Midkine Antibody (1011522) [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human Midkine
  Lys23-Asp143
  Accession # P21741
• Specificity: Detects human Midkine in direct ELISAs.
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-25 ug/mL
  • Immunohistochemistry 5-25 ug/mL
  • Immunoprecipitation 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Midkine Antibody (1011522) [Unconjugated]

• 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

Background

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.

1. Bohlen, P. and I. Kovesdi (1991) Prog. Growth Factor Res. 3:143.
2. Muramatsu, T. (1993) Int. J. Dev. Biol. 37:183.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Midkine Antibody (MAB2582)(1)

Publication using MAB2582

• Ayoubi, R;Southern, K;Laflamme, C;NeuroSGC/YCharOS Collaborative Group, ; A guide to selecting high-performing antibodies for human Midkine for use in Western blot and immunoprecipitation F1000Research 2024-08-01 [PMID: 39092005] (Immunoprecipitation, Western Blot, Human)

Bioinformatics

• Uniprot:
  • P21741()