CSN7b Overexpression Lysate

Images

 
Western Blot: CSN7b Overexpression Lysate (Native) [NBP2-06417] - CSN7b Overexpression Lysate (Native) Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for COPS7B.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

CSN7b Overexpression Lysate Summary

Description

CSN7b Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC221465

Accession#: NM_022730

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
COPS7B

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
29.4 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.

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 CSN7b Overexpression Lysate

  • Arabidopsis, homolog) subunit 7B
  • COP9 constitutive photomorphogenic homolog subunit 7B (Arabidopsis)
  • COP9 signalosome complex subunit 7b
  • CSN7BMGC111077
  • FLJ12612
  • Signalosome subunit 7b

Background

Component of the COP9 signalosome complex (CSN), is a complex involved in various cellular and developmental processes. The CSN complex is an essential regulator of the ubiquitin (Ubl) conjugation pathway by mediating the deneddylation of the cullin subunits of SCF type E3 ligase complexes, leading to decrease the Ubl ligase activity of SCF type complexes such as SCF, CSA or DDB2. The complex is also involved in phosphorylation of p53/TP53, c-jun/JUN, IkappaBalpha/NFKBIA, ITPK1 and IRF8/ICSBP, possibly via its association with CK2 and PKD kinases. CSN dependent phosphorylation of TP53 and JUN promotes and protects degradation by the Ubl system, respectively.

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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Product General Protocols

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Video Protocols

WB Video Protocol

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Bioinformatics

Gene Symbol COPS7B