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AB266247

Human PCNP knockout HEK-293T cell line

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PCNP KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 13 bp deletion in exon 1 and 1 bp insertion in exon 1.

View Alternative Names

AI647035, Ab2 416, PCNP_HUMAN, PEST proteolytic signal-containing nuclear protein, PEST-containing nuclear protein

2 Images
Sanger Sequencing - Human PCNP knockout HEK-293T cell line (AB266247)
  • Sanger seq

Unknown

Sanger Sequencing - Human PCNP knockout HEK-293T cell line (AB266247)

Allele-1 : 13 bp deletion in exon1

Sanger Sequencing - Human PCNP knockout HEK-293T cell line (AB266247)
  • Sanger seq

Unknown

Sanger Sequencing - Human PCNP knockout HEK-293T cell line (AB266247)

Allele-2 : 1 bp insertion in exon 1.

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, 13 bp deletion in exon 1 and 1 bp insertion in exon 1

Product details

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, 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?

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Properties and storage information

Gene name
PCNP
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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.

PCNP also called PEST-containing nuclear protein consists of around 178 amino acids and has a mass of approximately 21 kDa. PCNP localizes mainly in the nucleus of cells and shows expression in various tissues including the testis and skeletal muscle. The presence of PEST sequences suggests the target may experience rapid degradation which is typical for proteins that regulate essential cellular processes.
Biological function summary

This protein participates in cell cycle regulation and ubiquitination pathways suggesting its role in protein turnover within cells. PCNP often interacts with other proteins as part of larger complexes to influence proteolysis and cell cycle progression. The interaction with molecules containing ubiquitin-like domains indicates its involvement in tagging specific proteins for degradation helping maintain cellular homeostasis.

Pathways

PCNP connects strongly to the ubiquitin-proteasome pathway and the regulation of transcription. Its interactions with proteins such as RNF115 indicate its role in protein ubiquitination a fundamental process for maintaining cell integrity and function. Another relevant link is to cyclin-dependent kinases suggesting a contribution to cell cycle checkpoints and transitions essential for controlled cell division and proliferation.

PCNP's role highlights its potential involvement in oncogenesis and neurodegenerative disorders. Aberrant expression or regulation can connect PCNP to cancer progression by influencing the degradation of cell cycle regulators. Additionally defects in ubiquitination processes can tie this protein to conditions such as Alzheimer's disease where misfolded proteins accumulate. Interaction with proteins like beta-amyloid precursor protein suggests a possible link to neurodegenerative pathways impacting both disease development and progression.

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 protocols

Product promise

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