Midkine Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Midkine (MK) is a 13 kDa heparin-binding polypeptide which enhances neurite outgrowth, neuronal cell survival and plasminogen activator activity. MK is structurally divided into two domains, and most of the biological activities are located on the C-terminal domain. Both domains consist of three antiparallel beta-strands, but the C-terminal domain has a long flexible hairpin loop where a heparin-binding consensus sequence is located (1). MK is a member of a highly conserved, developmentally regulated human gene family. The gene product exhibits neurite outgrowth-promoting activity and may play a role in nervous system development and/or maintenance. Expression of MK is predominant only for a short period from approximately one-half to two-thirds of the way through gestation; before and after that, it is barely detectable (2)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Array NBP2-11177

• Midkine Antibodies
• Midkine ELISA Kit
• Midkine Lysates
• Midkine Proteins
• Midkine ELISA
• Midkine Proteins and Enzymes