Pleiotrophin/PTN in PC‑3 Human Cell Line.
Pleiotrophin/PTN was detected in immersion fixed PC‑3 human prostate cancer cell line using Rat Anti-Human Pleiotrophin/PTN Monoclonal Antibody (Catalog # MAB252) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
Reconstitution Buffer Available
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.
Pleiotrophin (PTN), also called heparin-binding growth-associated molecule (HB-GAM), heparin-binding neurotrophic factor (HBNF), heparin-affinity regulatory peptide (HARP), or osteoblast-specific factor (OSF-1), is an 18 kDa secreted, strongly heparin-binding, developmentally regulated cytokine (1 ‑ 3). PTN and Midkine share 50% amino acid (aa) sequence identity, share some functions, and constitute a family (1 ‑ 3). The second of two TSP1 domains contains the highest affinity binding site for heparin (4, 5). A 15 kDa form which lacks the C-terminus is mitogenic for glioblastoma cells, while full-length PTN is not (6). PTN is a highly conserved protein; human, mouse, rat, canine, porcine, equine and bovine PTN share 98% aa sequence identity or greater. During development, PTN is involved in development of brain, bone, and organs undergoing branching morphogenesis (3). In the adult, it is induced by PDGF and upregulated in many cancers, hematopoietic stem cells and tissues undergoing remodeling (7 ‑ 10). Cell surface receptors for PTN include Syndecan-3 (which mediates neurite outgrowth) and the receptor tyrosine phosphatase PTPRB, also called RPTP beta / zeta (3, 11 ‑ 13). Heparin binding is necessary for engaging these receptors (7, 8). PTN causes PTPRB dimerization and inactivates its phosphatase activity, which allows increased tyrosine phosphorylation of its substrates (12 ‑ 14). One such substrate is the WNT pathway molecule beta -catenin, allowing crosstalk of PTN with WNTs (12). PTN activation of the receptor ALK (anaplastic lymphoma kinase) is indirect through PTPRB, and mediates mitogenic, transforming and angiogenic activities of PTN (2, 5, 6, 13). Increased expression of PTN is correlated with neuronal development or stresses such as brain ischemia and Parkinson’s disease (2, 3, 7, 8). Both PTN and Midkine have demonstrated bactericidal activity, but only in the absence of heparin (15).
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