IKK beta Overexpression Lysate

Name: IKK beta Overexpression Lysate
Brand: Novus Biologicals
SKU: NBP2-10983

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-10983

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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 IKK beta Overexpression Lysate

IKK beta Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC219154

Accession#: NM_001556

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

86.4 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 IKK beta Overexpression Lysate

Western Blot: IKK beta Overexpression Lysate [NBP2-10983]

Western Blot: IKK beta Overexpression Lysate (Adult Normal) [NBP2-10983] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for IKK beta.

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: IKK beta

NF-kB (nuclear factor kB) is sequestered in the cytoplasm by IkB family of inhibitory proteins that mask the nuclear localization signal of NF-kB thereby preventing translocation of NF-kB to the nucleus (1). External stimuli such as tumor necrosis factor or other cytokines results in phosphorylation and degradation of IkB releasing NF-kB dimers. NF-kB dimer subsequently translocates to the nucleus and activates target genes. Synthesis of IkBa is autoregulated (2). IkB proteins are phosphorylated by IkB kinase complex consisting of at least three proteins, IKK1/a, IKK2/b, and IKK3/g (3-5). IKK1/a and IKK2/b are phosphorylated by NF-kB-inducing kinase (NIK) (6) and MAP kinase kinase kinase-1 (MEKK1) (7), respectively. Recent studies have shown that IKK2/b, not IKK1/a, is the target proinflammatory stimuli (9). Targeted disruption of IKK2/b gene in mice also results in extensive liver damage from apotopsis and death as embryo (9).

Long Name

IkB Kinase beta

Alternate Names

IkBKB, IKK2, NFKBIKB

Gene Symbol

IKBKB

Additional IKK beta Products

Product Documents for IKK beta Overexpression Lysate

Product Datasheets

Download Product Datasheets

COA

SDS

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Product Specific Notices for IKK beta 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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