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LIM kinase 2 Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-06997

Novus Biologicals, part of Bio-Techne
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NBP2-06997

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for LIM kinase 2 Overexpression Lysate

LIM kinase 2 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC220400

Accession#: NM_016733

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

69.7 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 LIM kinase 2 Overexpression Lysate

Western Blot: LIM kinase 2 Overexpression Lysate [NBP2-06997]

Western Blot: LIM kinase 2 Overexpression Lysate [NBP2-06997]

Western Blot: LIM kinase 2 Overexpression Lysate (Adult Normal) [NBP2-06997] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for LIM kinase 2.

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: LIM kinase 2

LIM-kinase 2 (LIMK2), a 75-kDa member of the LIM kinase family, is responsible for regulating actin polymerization via phosphorylation and inactivation of the actin depolymerizing factors ADF/cofilin (1). LIMK2 protein is found in the cytoplasm during interphase, redistributed to the mitotic spindle and then to the spindle midzone during anaphase to telophase (2). It has been shown that ROCK, a downstream protein kinase of Rho, activates LIMK2 by directly phosphorylates threonine 505 residue. Since activity of LIMK2 is cell cycle dependant, LIMK2 activity is markedly increased when HeLa cells are synchronized at mitosis with nocodazole treatment (3).

Long Name

LIM domain kinase 2

Alternate Names

LIMK-2, LIMK2

Gene Symbol

LIMK2

Additional LIM kinase 2 Products

Product Documents for LIM kinase 2 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 LIM kinase 2 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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