UBPY/USP8 Overexpression Lysate

Name: UBPY/USP8 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-17672

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-17672

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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 UBPY/USP8 Overexpression Lysate

UBPY/USP8 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC216926

Accession#: NM_005154

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

127.3 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 UBPY/USP8 Overexpression Lysate

Western Blot: UBPY/USP8 Overexpression Lysate [NBL1-17672]

Western Blot: UBPY/USP8 Overexpression Lysate (Adult Normal) [NBL1-17672] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for UBPY/USP8.

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: USP8

Ubiquitin Specific Peptidase 8 (USP8), also known as Ubiquitin Isopeptidase Y (UBPY), is a widely expressed deubiquitinating enzyme belonging to the peptidase C19 family. It has a predicted molecular weight of 127.5 kDa. Human USP8 is 1118 amino acids (aa) in length and shares 84% aa sequence identity with the mouse and rat orthologs. It contains an N-terminal MIT domain (aa 33-116) that mediates endosomal localization, CHMP-binding, and maintenance of ESCRT-0. USP8 also has a rhodanese domain (aa 181-319) that binds NRDP1 Ubiquitin ligase (E3), a SH3 domain binding sequence (aa 405-413), and a C-terminal catalytic domain (aa 734-1110). USP8 is a growth-regulated enzyme that controls the internalization and endocytic trafficking of cell surface receptors. Some receptors are targeted for internalization and degradation by ubiquitination. USP8 has been shown to disrupt the down-regulation of multiple receptors, including EGF R/ErbB1, ErbB2/Her2, and Smoothened, via their deubiquitination. Conversely, USP8 appears to have the opposite effect on the trafficking of CXCR4, PAR2, and the delta-opioid receptor. Depletion or catalytic inactivation of USP8 stabilized their expression. It is thought that deubiquitination of these receptors down-stream of the sorting endosome commits them to lysosomal degradation. USP8 can be phosphorylated at Ser680 allowing for 14-3-3-epsilon binding, which subsequently inhibits USP8 activity. Additionally, USP8 undergoes tyrosine phosphorylation at its N-terminus following EGF activation of the EGF R/ErbB1 / ErbB2/Her2 receptor complex.

Long Name

Ubiquitin Specific Protease 8

Alternate Names

UBPY

Gene Symbol

USP8

Additional USP8 Products

Product Documents for UBPY/USP8 Overexpression Lysate

Product Datasheets

Download Product Datasheets

COA

SDS

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Product Specific Notices for UBPY/USP8 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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