Key features and details
- One-wash 90 minute protocol
- Sensitivity: 14.4 pg/ml
- Range: 39.1 pg/ml - 2500 pg/ml
- Sample type: Cell culture extracts, Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Serum
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Product nameHuman Midkine ELISA Kit
See all Midkine kits
Intra-assay Sample n Mean SD CV% HepG2 8 2% Inter-assay Sample n Mean SD CV% HepG2 3 3.9%
Sample typeCell culture supernatant, Serum, Cell culture extracts, Hep Plasma, EDTA Plasma, Cit plasma
Assay typeSandwich (quantitative)
Range39.1 pg/ml - 2500 pg/ml
Sample specific recovery Sample type Average % Range Cell culture supernatant 100 82% - 115% Serum 114 113% - 115% Cell culture extracts 156 152% - 160% Hep Plasma 83 73% - 90% EDTA Plasma 112 109% - 114% Cit plasma 128 115% - 137%
Assay time1h 30m
Assay durationOne step assay
Species reactivityReacts with: Human
Does not react with: Cow, Pig
Human Midkine ELISA Kit (ab193761) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Midkine protein in cit plasma, edta plasma, hep plasma, cell culture extracts, serum, and cell culture supernatant. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human Midkine with 14.4 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
Midkine (MK), also known as neurite growth-promoting factor 2 (NEGF2), is a 13kDa heparin-binding growth factor or cytokine composed of two domains: The N-terminally located N-domain and the C-terminally located C-domain, which are connected by a hinge. The C-domain plays a role in neuronal development, whereas the N-domain is important for protein stability and dimerization. MK is involved in development, reproduction, repair, inflammation, innate immunity, control of blood pressure and angiogenesis.
MK is strongly expressed during embryogenesis and due to its distribution in the embryo it has been proposed to play a role in neurogenesis, epithelial-mesenchymal interactions and mesoderm remodeling. Expression in adult tissues is restricted to the kidney, epidermis, bronchial epithelium, lymphocytes and macrophages, but it is strongly expressed in the brain, kidney, blood vessels and heart after tissue injury as well as during inflammation, infection and oncogenesis. During inflammation, substratum-bound MK enhances neutrophil and macrophage migration directly and through induction of chemokine expression. On the other hand, soluble MK is associated with differentiation of regulatory T-cells, induction of epithelial-mesenchymal transition, angiogenesis, fibrinolytic and anti-microbial activity.
MK signaling is mediated by cell surface receptors as well as membrane proteins such as Protein Tyrosine Phosphatase ζ (PTPζ), low density lipoprotein receptor-related proteins (LRP), Notch2, integrins, anaplastic lymphoma kinase (ALK) and neuroglycan C. Binding of MK to PTPζ induces tyrosine phosphorylation in β-catenin and Wnt signaling inhibition. Furthermore it induces phosphorylation of PI3K, MAPK, PKC and Src family kinase. Binding of MK to LRP leads to embryonic neuronal survival and prevention of hypoxic injury via Akt and HIF1α. Binding of MK to integrins activates focal adhesion kinase, paxillin and STAT1α pathway leading to increase invasiveness of cancer cells. Activation of ALK by MK leads to phosphorylation of IRS-1, MAPK, PI3K and activation of NF-κB.
Due to its multifunctionality, MK has become an emerging target of drug development for the treatment of multiple diseases. On the one hand, administration of MK ameliorates ischemic injury, enhances oocyte maturation and promotes neurogenesis therefore limiting the progression of neurodegenerative diseases. However on the other hand, due to its over-expression in malignant tumors and in inflammation, MK inhibitors may be useful in the treatment of cancer, multiple sclerosis, hypertension and osteoporosis.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
PlatformMicroplate (12 x 8 well strips)
Storage instructionsStore at +4°C. Please refer to protocols.
Components 1 x 96 tests 10X Human Midkine Capture Antibody 1 x 600µl 10X Human Midkine Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent CPI - HAMA Blocker (ab193969) 1 x 6ml Human Midkine Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 50ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml
FunctionDevelopmentally regulated, secreted growth factor homologous to pleiotrophin (PTN), which has heparin binding activity. Binds anaplastic lymphoma kinase (ALK) which induces ALK activation and subsequent phosphorylation of the insulin receptor substrate (IRS1), followed by the activation of mitogen-activated protein kinase (MAPK) and PI3-kinase, and the induction of cell proliferation. Involved in neointima formation after arterial injury, possibly by mediating leukocyte recruitment. Also involved in early fetal adrenal gland development.
Tissue specificityExpressed in various tumor cell lines. In insulinoma tissue predominantly expressed in precancerous lesions.
Sequence similaritiesBelongs to the pleiotrophin family.
- Information by UniProt
- Amphiregulin-associated protein
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
The Midkine standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
The concentrations of Midkine were measured in duplicates, interpolated from the Midkine standard curves and corrected for sample dilution. Undiluted samples are as follows: HepG2 cell culture supernatant 2.5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Midkine concentration was determined to be 6,242 pg/mL in serum.
The concentrations of Midkine were measured in duplicate and interpolated from the Midkine standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Midkine concentration was determined to be 2641 pg/mL in HepG2 cell extract and 1179 pg/mL in SH-SY5Y cell extract.
The concentrations of Midkine were measured in duplicates, interpolated from the Midkine standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (EDTA) 50%, plasma (citrate) 25%, and plasma (heparin) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Midkine concentration was determined to be 243 pg/mL in serum, 183 pg/mL in plasma (EDTA), 449 pg/mL plasma (citrate) and 303 pg/mL in plasma (heparin).
Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Midkine concentration was determined to be 164 pg/mL with a range of 69 – 594 pg/mL.
ab193761 has been referenced in 3 publications.
- Meng X et al. DNA damage repair alterations modulate M2 polarization of microglia to remodel the tumor microenvironment via the p53-mediated MDK expression in glioma. EBioMedicine 41:185-199 (2019). PubMed: 30773478
- Rice LM et al. Serum biomarker for diagnostic evaluation of pulmonary arterial hypertension in systemic sclerosis. Arthritis Res Ther 20:185 (2018). PubMed: 30115106
- Su Z et al. Circulating midkine in children with Henoch-Schönlein purpura: Clinical implications. Int Immunopharmacol 39:246-250 (2016). Sandwich ELISA ; Human . PubMed: 27497193