PKA C beta Overexpression Lysate

Name: PKA C beta Overexpression Lysate
Brand: Novus Biologicals
SKU: NBP2-04480

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-04480

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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 PKA C beta Overexpression Lysate

PKA C beta Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC202131

Accession#: NM_207578

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

29.5 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 PKA C beta Overexpression Lysate

Western Blot: PKA C beta Overexpression Lysate [NBP2-04480]

Western Blot: PKA C-beta Overexpression Lysate (Adult Normal) [NBP2-04480] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for PKA C-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: PKA C beta

The second messenger cyclic AMP (cAMP) mediates diverse cellular responses to external signals such as proliferation, ion transport, regulation of metabolism and gene transcription by activation of the cAMP-dependent protein kinase (cAPK or PKA). Activation of PKA occurs when cAMP binds to the two regulatory subunits of the tetrameric PKA holoenzyme resulting in release of active catalytic subunits. Three catalytic (C) subunits have been identified, designated Calpha, Cbeta and Cgamma, that each represent specific gene products. Calpha and Cbeta are closely related (93% amino acid sequence similarity), whereas Cgamma displays 83% and 79% similarity to Calpha and Cbeta, respectively. Activation of transcription upon elevation of cAMP levels results from translocation of PKA to the nucleus where it phosphorylates the transcription factor cAMP response element binding protein (CREB) on serine 133 which in turn leads to TFIIB binding to TATA-box-binding protein TBP1, thus linking phospho-CREB to the pol II transcription initiation complex.

Long Name

cAMP-dependent Protein Kinase Catalytic Subunit beta

Alternate Names

cAPKbeta, PRKACB

Gene Symbol

PRKACB

Additional PKA C beta Products

Product Documents for PKA C beta Overexpression Lysate

Product Datasheets

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COA

SDS

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Product Specific Notices for PKA C 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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