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AB267332

Human JOSD1 knockout HEK-293T cell line

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JOSD1 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion 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'.
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Sanger Sequencing - Human JOSD1 knockout HEK-293T cell line (AB267332)
  • Sanger seq

Unknown

Sanger Sequencing - Human JOSD1 knockout HEK-293T cell line (AB267332)

Homozygous : 1 bp insertion in exon1

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 insertion in exon 1

Product details

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
JOSD1
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.

JOSD1 also known as Josephin domain-containing protein 1 functions as a deubiquitinating enzyme. It specifically removes ubiquitin from protein substrates which regulates protein degradation localization and activity in cells. The protein mass is approximately 45 kDa. JOSD1 is expressed in various tissues with significant levels seen in the brain liver and kidney. By modulating the ubiquitination status of substrates JOSD1 influences cellular processes like signal transduction and response to stress.
Biological function summary

This protein modifies the stability and activity of other proteins by removing ubiquitin molecules. It acts as part of a larger deubiquitinase complex which interacts with protein networks to maintain cellular homeostasis. Through its enzyme activity JOSD1 contributes to the regulation of processes such as protein quality control and cell cycle progression.

Pathways

Scientists have placed JOSD1 in the ubiquitin-proteasome system an important pathway responsible for protein turnover in cells. It has connections with other proteins like USP7 and USP9X in this pathway reflecting its coordination role in protein deubiquitination. JOSD1 also participates in the endoplasmic reticulum-associated degradation (ERAD) pathway helping to eliminate misfolded proteins important for maintaining protein folding homeostasis within the cell.

Abnormal activity of JOSD1 can connect to neurodegenerative disorders such as Alzheimer's disease where protein misfolding and aggregation play significant roles. JOSD1's interaction with proteins like tau and amyloid precursor protein suggests its potential to influence disease progression. Additionally research associates JOSD1 with liver cancer where dysregulated ubiquitin signaling may contribute to tumor development and progression.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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For full details, please see our Terms & Conditions

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