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UCHL1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 45 bp deletion in exon 1.

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Images

Western blot - Human UCHL1 (PGP9.5) knockout HEK-293T cell line (AB255443), expandable thumbnail
  • Sanger Sequencing - Human UCHL1 (PGP9.5) knockout HEK-293T cell line (AB255443), expandable thumbnail

Key facts

Cell type
HEK-293T
Species or organism
Human
Tissue
Kidney
Form
Liquid
Knockout validation
Sanger Sequencing, Western blot
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 45 bp deletion in exon 1

Alternative names

Recommended products

UCHL1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 45 bp deletion in exon 1.

Key facts

Cell type
HEK-293T
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 45 bp deletion in exon 1
Concentration
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Properties

Gene name
UCHL1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous

Quality control

STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female

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.

Storage

Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

Notes

Recommended control: Human wild-type HEK293T cell line (ab255449). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

Protein Gene Product 9.5 (PGP9.5) also known as ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1) is an enzyme of approximately 27 kDa mass. PGP9.5 is highly expressed in neurons and neuroendocrine cells and is involved in the processing of ubiquitinated proteins. Ubiquitination and deubiquitination are critical for maintaining cellular protein homeostasis. PGP9.5 is present in the central and peripheral nervous systems. Researchers often utilize PGP9.5 as a neuronal marker due to its strong expression in nerve tissues. Antibody clone 13C4 is commonly used in PGP9.5 immunohistochemistry to visualize this protein in histopathology sections.

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.

Associated diseases and disorders

Defective PGP9.5 function associates mainly with neurodegenerative diseases such as Parkinson's disease and Alzheimer's disease. Mutations or dysregulation in PGP9.5 expression are implicated in the pathogenesis of these conditions affecting protein degradation and neuronal health. Other proteins like α-synuclein in Parkinson's disease interact with PGP9.5 pathways indicating their involvement in disease mechanisms. Researchers consider PGP9.5 a potential biomarker for neuronal pathology through methods like PGP9.5 IHC allowing for evaluation of nerve damage and neuroendocrine tumor diagnosis.

Product promise

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In the unlikely event of one of our products not working as expected, you are covered by our product promise.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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