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AB264736

Human ZMPSTE24 knockout HeLa cell line

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ZMPSTE24 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.

View Alternative Names

CAAX prenyl protease 1 homolog, FACE1_HUMAN, Farnesylated proteins-converting enzyme 1, Prenyl protein-specific endoprotease 1, STE24, Zinc metalloproteinase Ste24 homolog

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Sanger Sequencing - Human ZMPSTE24 knockout HeLa cell line (AB264736)
  • Sanger seq

Unknown

Sanger Sequencing - Human ZMPSTE24 knockout HeLa cell line (AB264736)

Homozygous : 1 bp insertion in exon 1.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1

Disease

Adenocarcinoma

Product details

Recommended control: Human wild-type HeLa cell line (ab255928). 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
ZMPSTE24
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Zygosity
Homozygous
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.

ZMPSTE24 also known as FACE1 or STE24 is a zinc metalloprotease enzyme involved in the post-translational modification of prelamin A to mature lamin A. This enzyme specifically cleaves the C-terminal tail of prelamin A a critical step required for proper nuclear envelope structure and function. ZMPSTE24 has a molecular mass of approximately 54 kDa and is expressed in several tissues including skeletal muscle heart and skin. Its activity primarily takes place in the endoplasmic reticulum membrane where it executes its proteolytic functions.
Biological function summary

The alteration of prelamin A to lamin A by ZMPSTE24 is necessary for maintaining nuclear shape and stability. Lamin A is a major component of the nuclear lamina providing structural support to the nuclear envelope. ZMPSTE24 is part of a proteolytic complex that ensures the proper maturation of lamin A which plays a significant role in DNA replication and cellular division. Deficiency in ZMPSTE24 activity can lead to the accumulation of farnesylated prelamin A disrupting nuclear architecture and cellular processes.

Pathways

The function of ZMPSTE24 integrates into the broader context of the lamin A maturation pathway significant for nuclear integrity. It collaborates with other enzymes like farnesyltransferase and icmt during the maturation process of prelamin A to lamin A. ZMPSTE24 also interacts with the protein lamin B which is an associate in the nuclear lamina. This cooperation within the pathway ensures the functional execution of nuclear mechanical properties and DNA repair processes.

The abnormal function or mutation of ZMPSTE24 connects direly with disorders such as restrictive dermopathy and Hutchinson-Gilford progeria syndrome (HGPS). In HGPS defective ZMPSTE24 fails to process prelamin A leading to the build-up of a toxic version of prelamin A termed progerin. This accumulation contributes to premature aging phenotypes. The disorder correlates with mutations in LMNA a gene encoding lamins A and C highlighting the tandem dependency between ZMPSTE24 and lamin A in maintaining nuclear structure and overall cellular health.

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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