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CDK8 Overexpression Lysate

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Western Blot: Cdk8 Overexpression Lysate (Adult Normal) [NBL1-09046] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Cdk8.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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CDK8 Overexpression Lysate Summary

Description

CDK8 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC212592

Accession#: NM_001260

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
CDK8

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
53.1 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 CDK8 Overexpression Lysate

  • CDK8 protein kinase
  • CDK8
  • Cell division protein kinase 8
  • cyclin-dependent kinase 8
  • EC 2.7.11
  • EC 2.7.11.22
  • EC 2.7.11.23
  • K35
  • Mediator complex subunit CDK8
  • Mediator of RNA polymerase II transcription subunit CDK8
  • MGC126074
  • MGC126075
  • Protein kinase K35

Background

Cdk8 is encoded by this gene is a member of the cyclin-dependent protein kinase (CDK) family. CDK family members are highly similar to the gene products of Saccharomyces cerevisiae cdc28, and Schizosaccharomyces pombe cdc2, and are known to be important regulators of cell cycle progression. This kinase and its regulatory subunit cyclin C are components of the RNA polymerase II holoenzyme complex, which phosphorylates the carboxy-terminal domain (CTD) of the largest subunit of RNA polymerase II. This kinase has also been shown to regulate transcription by targeting the CDK7/cyclin H subunits of the general transcription initiation factor IIH (TFIIH), thus providing a link between the 'Mediator-like' protein complexes and the basal transcription machinery.

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 CDK8