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OTUB1 KO cell line available to order. KO validated by Immunocytochemistry, Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 1 and Insertion of the selection cassette in exon 1.

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Images

Western blot - Human OTUB1 knockout HEK-293T cell line (AB266551), expandable thumbnail
  • Western blot - Human OTUB1 knockout HEK-293T cell line (AB266551), expandable thumbnail
  • Western blot - Human OTUB1 knockout HEK-293T cell line (AB266551), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Human OTUB1 knockout HEK-293T cell line (AB266551), expandable thumbnail
  • Cell Culture - Human OTUB1 knockout HEK-293T cell line (AB266551), expandable thumbnail

Key facts

Cell type
HEK-293T
Species or organism
Human
Tissue
Kidney
Form
Liquid
Knockout validation
Immunocytochemistry, Sanger Sequencing, Western blot
Mutation description
Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 1 and Insertion of the selection cassette in exon 1

Alternative names

Deubiquitinating enzyme OTUB1, HSPC263, OTB1, OTU domain, ubiquitin aldehyde binding 1, OTU domain-containing Ubal-binding protein 1, OTU domain-containing ubiquitin aldehyde-binding protein 1, OTU-domain Ubal-binding 1, OTUB1_HUMAN, Otubain-1, Ubiquitin thioesterase OTUB1, Ubiquitin-specific-processing protease OTUB1, hOTU1, ubiquitin-specific protease otubain 1

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OTUB1 KO cell line available to order. KO validated by Immunocytochemistry, Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 1 and Insertion of the selection cassette in exon 1.

Key facts

Cell type
HEK-293T
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 1 and Insertion of the selection cassette in exon 1
Concentration
Loading...

Properties

Gene name
OTUB1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Immunocytochemistry, Sanger Sequencing, Western blot

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, 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.

Supplementary info

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

The OTUB1 protein also known as OTU domain-containing ubiquitin aldehyde-binding protein 1 plays an important role in the removal of ubiquitin from proteins. This process is important for regulating protein stability and function within the cell. OTUB1 has a mass of approximately 32 kDa and is expressed ubiquitously across various tissues indicating its essential nature in cellular functions.

Biological function summary

OTUB1 functions as a deubiquitinating enzyme that cleaves ubiquitin from substrates influencing various cellular processes. It acts independently or as part of larger protein complexes. By controlling protein degradation pathways OTUB1 impacts protein turnover and cellular homeostasis. Interactions with other proteins allow OTUB1 to exert influence on different signaling pathways and cellular stress responses.

Pathways

OTUB1 holds a significant position in the regulation of the ubiquitin-proteasome system and DNA damage repair pathway. It closely interacts with proteins such as UBE2N affecting the DNA damage response. Additionally OTUB1 regulates processes through modulating pathways like NF-kB by interacting with and stabilizing specific protein factors involved.

Associated diseases and disorders

OTUB1’s function connects it to various pathological states like cancer and neurodegenerative diseases. Aberrant expression or mutations in OTUB1 can lead to disruptions in cell cycle regulation and apoptosis contributing to oncogenesis. In the context of neurodegenerative diseases OTUB1 is related to tau stabilization with proteins such as HSP90 playing a role in the same regulatory mechanisms highlighting its relevance in these disorders.

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Product protocols

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

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