Mark3 Overexpression Lysate

Name: Mark3 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-12899

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-12899

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for Mark3 Overexpression Lysate

Mark3 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC205758

Accession#: NM_002376

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

81.3 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 Mark3 Overexpression Lysate

Western Blot: Mark3 Overexpression Lysate [NBL1-12899]

Western Blot: Mark3 Overexpression Lysate (Adult Normal) [NBL1-12899] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Mark3.

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: Mark3

MARK3 is a dual-specificity protein kinase that controls entry into mitosis by dephosphorylating Cdc2 on both threonine 14 and tyrosine 15. MARK3 is phosphorylated on serine 216 throughout interphase but not during mitosis. Serine 216 phosphorylation mediates the binding of 14-3-3 protein to MARK3, and MARK3/14-3-3 complexes are present throughout interphase but not during mitosis (1). Kinase suppressor of Ras (KSR) is a conserved component of the Ras pathway that interacts directly with MEK and MAPK. It has been shown that KSR1 translocates from the cytoplasm to the cell surface in response to growth factor treatment and that this process is regulated by MARK3 (2). MARK3 seems to be a positive regulator of the beta-catenin pathway and an inhibitor of the JNK pathway. These findings show that MARK3, a regulator of polarity, is also a modulator of Wnt-beta-catenin signalling, indicating a link between two important developmental pathways (3).

Alternate Names

Cdc25C-associated protein kinase 1, cTAK1, C-TAK1, CTAK1ELKL motif kinase 2, EC 2.7.11, EC 2.7.11.1, EMK2, EMK-2, KP78, MAP/microtubule affinity-regulating kinase 3, PAR1A, Protein kinase STK10, Ser/Thr protein kinase PAR-1, Serine/threonine-protein kinase p78

Gene Symbol

MARK3

Additional Mark3 Products

Product Documents for Mark3 Overexpression Lysate

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Mark3 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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