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AB266121

Human PCNT (Pericentrin) knockout HEK-293T cell line

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PCNT KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 12. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
2 Images
Cell Culture - Human PCNT (Pericentrin) knockout HEK-293T cell line (AB266121)
  • Cell Culture

Unknown

Cell Culture - Human PCNT (Pericentrin) knockout HEK-293T cell line (AB266121)

Representative images of PCNT knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.

Sanger Sequencing - Human PCNT (Pericentrin) knockout HEK-293T cell line (AB266121)
  • Sanger seq

Unknown

Sanger Sequencing - Human PCNT (Pericentrin) knockout HEK-293T cell line (AB266121)

Homozygous : 1 bp deletion in exon 12

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, Homozygous: 1 bp deletion in exon 12

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?

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Properties and storage information

Gene name
PCNT
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.

Pericentrin also known as PCNT is a large protein with a mass of approximately 360 kDa. It primarily localizes at the centrosome an organelle serving as the main microtubule organizing center in cells. Expressed in most cell types pericentrin is an essential structural component of the centrosome. Researchers often use pericentrin staining techniques to visualize centrosomes aiding in the study of cell division. This protein functions prominently as a centrosome marker facilitating centrosome image and imaging of centrosome-related structures.
Biological function summary

Pericentrin contributes significantly to maintaining centrosome integrity and organizing microtubules during cell division. It is a part of the pericentriolar material a complex that stabilizes and anchors microtubules to the centrosome. By forming a scaffold at the centrosome pericentrin plays an important role in recruiting other proteins like γ-tubulin enhancing its function in nucleating microtubules. The pericentrin protein is vital for proper spindle organization during mitosis affecting accurate chromosome segregation.

Pathways

Pericentrin is integral to the cell cycle and mitotic spindle assembly pathways. It interacts with several other proteins involved in mitosis including γ-tubulin and ninein to manage the assembly and stability of the mitotic spindle. By participating in these pathways pericentrin ensures correct cellular replication and division therefore maintaining genetic stability within organisms.

Mutations in the pericentrin gene have been linked to microcephalic osteodysplastic primordial dwarfism type II and Seckel syndrome. These conditions suggest a connection between defective pericentrin function and disruptions in cell division and growth. Studies also show that alterations in pericentrin levels can influence tumorigenesis by affecting centrosome function and chromosome segregation increasing the importance of this protein in cancer research.

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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