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TRAF6 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.

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Images

Western blot - Human TRAF6 knockout HeLa cell line (AB266009), expandable thumbnail
  • Western blot - Human TRAF6 knockout HeLa cell line (AB266009), expandable thumbnail
  • Sanger Sequencing - Human TRAF6 knockout HeLa cell line (AB266009), expandable thumbnail

Key facts

Cell type
HeLa
Species or organism
Human
Tissue
Cervix
Form
Liquid
Knockout validation
Sanger Sequencing, Western blot
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2

Alternative names

E3 ubiquitin-protein ligase TRAF6, Interleukin-1 signal transducer, MGC 3310, MGC:3310, OTTHUMP00000232772, OTTHUMP00000232773, RING finger protein 85, RNF 85, TNF receptor-associated factor 6, TNF receptor-associated factor 6, E3 ubiquitin protein ligase, TRAF6_HUMAN

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TRAF6 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.

Key facts

Cell type
HeLa
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2
Antibiotic resistance
Puromycin 1µg/mL
Disease
Adenocarcinoma
Concentration
Loading...

Properties

Gene name
TRAF6
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous

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 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.

Supplementary info

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

TRAF6 also known as TNF receptor-associated factor 6 is an E3 ubiquitin ligase that is part of the TRAF family. It has a molecular mass of approximately 60 kDa. TRAF6 is ubiquitously expressed in many types of tissues and cells including immune cells like macrophages and dendritic cells. Its primary role is to mediate signal transduction from members of the TNF receptor and Toll-like receptor (TLR) families which it achieves through polyubiquitination of target proteins. This process facilitates the recruitment of downstream signaling components necessary for cellular responses.

Biological function summary

TRAF6 plays a critical role in regulating immune and inflammatory responses. It acts within a complex often associating with other signaling proteins such as IRAK1 and TAK1 to activate downstream kinases like IKK and JNK. These signaling pathways lead to the activation of transcription factors such as NF-kB and AP-1 which regulate the expression of genes involved in inflammation and immune responses. TRAF6 is an important modulator of adaptive and innate immunity influencing both the maturation of immune cells and the body's response to infection.

Pathways

TRAF6 significantly contributes to the NF-kB and MAPK signaling pathways. In the NF-kB pathway TRAF6 mediates signal transduction following TLR engagement interacting with proteins such as MyD88 and RIP1 to drive inflammatory cytokine production. In the MAPK pathway it associates predominantly with TAK1 to facilitate the phosphorylation and activation of downstream kinases. These pathways are essential for transmitting signals from cell surface receptors to the nucleus controlling the magnitude of immune signaling and cellular stress responses.

Associated diseases and disorders

TRAF6 has a significant association with autoimmune diseases and certain cancers. In autoimmune disorders aberrant activation of TRAF6 can lead to prolonged inflammation and detrimental immune responses. For cancers TRAF6 is linked to tumor progression and metastasis acting alongside proteins like IRAK1 and RIP1 which can further promote malignancy through chronic inflammation and immune evasion. Understanding TRAF6's role in these diseases provides insights that could lead to targeted therapeutic strategies.

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

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