Human UCHL1 (PGP9.5) knockout HEK-293T cell lysate
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- WB
Unknown
Western blot - Human UCHL1 (PGP9.5) knockout HEK-293T cell lysate (AB263773)
Lane 1 : Wild-type Hap1 cell lysate (20 μg)
Lane 2 : UCHL1 knockout Hap1 cell lysate (20 μg)
Lane 3 : SH-SY5Y cell lysate (20 μg)
Lane 4 : Wild-type HEK-293T cell lysate (20 μg)
Lane 5 : UCHL1 knockout HEK-293T cell lysate (20 μg)
Lanes 1 - 5 : Merged signal (red and green). Green - ab108986 observed at 25 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab108986 was shown to react with PGP9.5 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab255443 (knockout cell lysate ab263773) was used. Wild-type and PGP9.5 knockout samples were subjected to SDS-PAGE. ab108986 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively.Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (<a href='/en-us/products/primary-antibodies/pgp95-antibody-epr4118-neuronal-marker-ab108986'>ab108986</a>) at 1/1000 dilution
Lane 1:
Wild-type HAP1 cell lysate at 20 µg
Lane 2:
UCHL1 knockout HAP1 cell lysate at 20 µg
Lane 2:
Western blot - Human UCHL1 (PGP9.5) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-uchl1-pgp95-knockout-hek-293t-cell-line-ab255443'>ab255443</a>)
Lane 3:
SH-SY5Y cell lysate at 20 µg
Lane 4:
Wild-type HEK-293T cell lysate at 20 µg
Lane 5:
UCHL1 knockout HEK-293T cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Predicted band size: 24 kDa
Observed band size: 25 kDa,37 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human UCHL1 (PGP9.5) knockout HEK-293T cell lysate (AB263773)
Homozygous : 45 bp deletion in exon 1
Complementary Products
Primary Antibodies
AB108986
Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker
BOND RX™ Validated|KO Validated|RabMAb|Recombinant|Lab Essentials|20ul selling size
![Western blot - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (AB108986)](https://content.abcam.com/products/images/pgp95-antibody-epr4118-neuronal-marker-ab108986--western-blot-img5852.jpg)
- 5
- 12
Reactivity data
Product details
Knockout cell lysate achieved by CRISPR/Cas9.
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 and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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
PGP9.5 plays a significant role in the ubiquitin-proteasome pathway where it regulates protein degradation. It functions by cleaving ubiquitin from ubiquitin-protein conjugates which can recycle ubiquitin for reuse impacting protein turnover. This process is important for removing misfolded or damaged proteins maintaining cellular integrity. PGP9.5 has not been documented as a part of any large multimeric complex; its action is rather individual yet vital in cellular processes.
Pathways
PGP9.5 is an important participant in the ubiquitin-dependent proteolysis pathway. This pathway controls the degradation of proteins that regulate cell cycle differentiation and neuromodulation. PGP9.5 works in tandem with other proteins such as the proteasome complex to remove proteins tagged for destruction. Another important pathway linked with PGP9.5 is the neuronal development pathway in which its activity supports neuron growth and repair.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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