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Midkine Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-10999

Novus Biologicals, part of Bio-Techne
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NBP2-10999

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for Midkine Overexpression Lysate

Midkine Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC221818

Accession#: NM_001012334

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.

Product Specifications

Application Notes

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

TMW

15.4 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Type

Overexpression

Scientific Data Images for Midkine Overexpression Lysate

Western Blot: Midkine Overexpression Lysate [NBP2-10999]

Western Blot: Midkine Overexpression Lysate [NBP2-10999]

Western Blot: Midkine Overexpression Lysate (Adult Normal) [NBP2-10999] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Midkine.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: Midkine

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)

Alternate Names

ARAP, MDK, MEK, MK1, NEGF2

Gene Symbol

MDK

Additional Midkine Products

Product Documents for Midkine Overexpression Lysate

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for 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.

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.

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