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PKA C beta Overexpression Lysate

Images

 
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.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

PKA C beta Overexpression Lysate Summary

Description

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.
Gene
PRKACB

Applications/Dilutions

Dilutions
  • Western Blot
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.
Theoretical MW
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.

Packaging, Storage & Formulations

Storage
Store at -80C. Avoid freeze-thaw cycles.
Buffer
RIPA buffer

Lysate Details for Array

Type
Overexpression

Notes

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.

Alternate Names for PKA C beta Overexpression Lysate

  • cAMP-dependent protein kinase catalytic beta subunit isoform 4ab
  • cAMP-dependent protein kinase catalytic subunit beta
  • cAPKbeta
  • DKFZp781I2452
  • EC 2.7.11
  • EC 2.7.11.11
  • MGC41879
  • MGC9320
  • PKA C beta
  • PKA C-beta
  • PKACb
  • PRKACB
  • protein kinase A catalytic subunit beta
  • protein kinase, cAMP-dependent, catalytic, beta

Background

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.

Limitations

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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Bioinformatics

Gene Symbol PRKACB