Human UCHL1 (PGP9.5) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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UCHL1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 45 bp deletion in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
Epididymis luminal protein 117, Epididymis secretory protein Li 53, HEL 117, HEL S 53, NDGOA, Neuron cytoplasmic protein 9.5, OTTHUMP00000218137, OTTHUMP00000218139, OTTHUMP00000218140, OTTHUMP00000218141, PGP 9.5, PGP95, Park 5, Protein gene product 9.5, UCHL1_HUMAN, Ubiquitin C terminal esterase L1, Ubiquitin C terminal hydrolase, Ubiquitin C terminal hydrolase L1, Ubiquitin carboxyl terminal esterase L1, Ubiquitin carboxyl-terminal hydrolase isozyme L1, Ubiquitin thioesterase L1, Ubiquitin thiolesterase, Ubiquitin thiolesterase L1
- WB
Unknown
Western blot - Human UCHL1 (PGP9.5) knockout HEK-293T cell line (AB255443)
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 (ab255443)
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 line (AB255443)
Homozygous : 45 bp deletion in exon 1.
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
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
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
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
Target data
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com