Human USP7 Baculovirus-Insect cells Overexpression Lysate

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Human USP7 Baculovirus-Insect cells Overexpression Lysate: Product Information

Product Description
This Human USP7 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of USP7 protein (Cat: 11681-H20B1) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect cells
Species
Human
Sequence Information
A DNA sequence encoding full length of human USP7 (NP_003461.2) (Met 1-Asn 1102) was fused with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
Molecule Mass
The recombinant human USP7/GST chimera consists of 1339 amino acids and has a calculated molecular mass of 156 kDa. It migrates as an approximately 140 band in SDS-PAGE under reducing conditions.

Human USP7 Baculovirus-Insect cells Overexpression Lysate: Usage Guide

Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Lysis Buffer
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Recommend Usage
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human USP7 Baculovirus-Insect cells Overexpression Lysate: Alternative Names

Human HAUSP Overexpression Lysate; Human TEF1 Overexpression Lysate

USP7 Background Information

Ubiquitin carboxyl-terminal hydrolase 7, also known as Ubiquitin thioesterase 7, Herpesvirus-associated ubiquitin-specific protease, Ubiquitin-specific-processing protease 7, USP7 and HAUSP, is a widely expressed protein which belongs to thepeptidase C19 family. USP7 is a member of the family of deubiquitinating enzymes. It is involved in the regulation of stress response pathways, epigenetic silencing and the progress of infections by DNA viruses. USP7 is a protein with a cysteine peptidase core, N- and C-terminal domains required for protein-protein interactions. USP7 contributes to epigenetic silencing of homeotic genes by Polycomb (Pc). USP7 cleaves ubiquitin fusion protein substrates. It deubiquitinates TP53/p53 and MDM2 and strongly stabilizes TP53 even in the presence of excess MDM2. USP7 also induces TP53-dependent cell growth repression and apoptosis. USP7 has key roles in the p53 pathway whereby it stabilizes both p53 and MDM2. Herpes simplex virus type 1 (HSV-1) regulatory protein ICP stimulates lytic infection and the reactivation of quiescent viral genomes. ICP interacts very strongly with USP7. USP7-mediated stabilization of ICP is dominant over ICP-induced degradation of USP7 during productive HSV-1 infection. The biological significance of the ICP-USP7 interaction may be most pronounced in natural infection situations, in which limited amounts of ICP are expressed.
Full Name
ubiquitin specific peptidase 7 (herpes virus-associated)
References
  • Zakut-Houri R., et al.,(1985), Human p53 cellular tumor antigen: cDNA sequence and expression in COS cells. EMBO J. 4:1251-1255.
  • Lamb P., et al., (1986), Characterization of the human p53 gene.Mol. Cell. Biol. 6:1379-1385.
  • Harlow E., et al.,(1985), Molecular cloning and in vitro expression of a cDNA clone for human cellular tumor antigen p53.Mol. Cell. Biol. 5:1601-1610.
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