Human USP14 (TGT) knockout HeLa cell lysate
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USP14 KO cell lysate available now. KO validated. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.
View Alternative Names
Deubiquitinating enzyme 14, UBP14_HUMAN, USP 14, Ubiquitin carboxyl-terminal hydrolase 14, Ubiquitin specific peptidase 14, Ubiquitin specific protease 14, Ubiquitin thiolesterase 14, Ubiquitin-specific-processing protease 14, tRNA guanine transglycosylase 60 kD subunit, tRNA-guanine transglycosylase
- WB
Lab
Western blot - Human USP14 (TGT) knockout HeLa cell lysate (AB257787)
Lane 1 : Wild-type HeLa cell lysate 20 μg
Lane 2 : USP14 knockout HeLa cell lysate 20 μg
Lane 3 : USP14 knockout HeLa cell lysate 20 μg
False colour image of Western blot : Anti-USP14/TGT antibody [EPR15943] - C-terminal staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab192618 was shown to bind specifically to USP14/TGT. A band was observed at 63 kDa in wild-type HeLa cell lysates with no signal observed at this size in USP14 knockout cell line ab266854 (knockout cell lysate ab257787). The band observed in the knockout lysate lane below 63 kDa is likely to represent a truncated form of USP14/TGT. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and USP14 knockout HeLa cell lysates were analysed.First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-USP14/TGT antibody [EPR15943] - C-terminal (<a href='/en-us/products/primary-antibodies/usp14-tgt-antibody-epr15943-c-terminal-ab192618'>ab192618</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lanes 2 - 3:
USP14 knockout HeLa cell lysate at 20 µg
Predicted band size: 56 kDa
Observed band size: 63 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human USP14 (TGT) knockout HeLa cell lysate (AB257787)
Homozygous : 1 bp insertion in exon 1
Reactivity data
Product details
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
USP14 functions in regulating protein turnover by removing ubiquitin from substrate proteins impacting their stability and degradation. USP14 is part of a larger protein complex associated with the proteasome specifically binding to the 19S regulatory particle. It is important in maintaining cellular protein homeostasis by editing ubiquitin chains on target proteins prior to degradation.
Pathways
USP14 participates in ubiquitin-proteasome pathways and ER-associated degradation pathway. In these processes it works closely with other proteasome proteins such as Rpn11 which also removes ubiquitin from proteins. The regulation of these pathways by USP14 directly affects protein quality control in cells influencing processes like cell cycle and stress response.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
![Flow Cytometry (Intracellular) - Anti-USP14/TGT antibody [EPR15943] - C-terminal (AB192618)](https://content.abcam.com/products/images/usp14-tgt-antibody-epr15943-c-terminal-ab192618--flow-cytometry-intracellular-img118085.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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