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AB257760

Human TRAF6 knockout HeLa cell lysate

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TRAF6 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2.
3 Images
Western blot - Human TRAF6 knockout HeLa cell lysate (AB257760)
  • WB

Lab

Western blot - Human TRAF6 knockout HeLa cell lysate (AB257760)

Lane 1 : Wild-type HeLa cell lysate (20μg)
Lane 2 : TRAF6 knockout HeLa cell lysate (20μg)
Lane 3 : HAP1 cell lysate (20μg)
Lane 4 : Daudi cell lysate (20μg)
Lanes 1- 4 : Merged signal (red and green). Green - ab40675 observed at 65 kDa. Red - loading control ab8245 observed at 37 kDa.
ab40675 Rabbit monoclonal [EP592Y] to TRAF6 was shown to specifically react with TRAF6 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab266009 (knockout cell lysate ab257760) was used. Wild-type and TRAF6 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab40675 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 μg/ml and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-TRAF6 antibody [EP592Y] (<a href='/en-us/products/primary-antibodies/traf6-antibody-ep592y-ab40675'>ab40675</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

TRAF6 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human TRAF6 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-traf6-knockout-hela-cell-line-ab266009'>ab266009</a>)

Lane 3:

HAP1 cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Predicted band size: 60 kDa

Observed band size: 65 kDa

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Western blot - Human TRAF6 knockout HeLa cell lysate (AB257760)
  • WB

Lab

Western blot - Human TRAF6 knockout HeLa cell lysate (AB257760)

Lane 1 : Wild-type HeLa cell lysate (20µg)

Lane 2 : TRAF6 knockout HeLa cell lysate (20µg)

Lane 3 : HAP1 cell lysate (20µg)

Lane 4 : Daudi cell lysate (20µg)

Lanes 1- 4 : Merged signal (red and green). Green - ab33915 observed at 65 kDa. Red - loading control ab8245 observed at 37 kDa.

ab33915 Rabbit monoclonal [EP591Y] to TRAF6 was shown to specifically react with TRAF6 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab266009 (knockout cell lysate ab257760) was used. Wild-type and TRAF6 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab33915 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 2000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-TRAF6 antibody [EP591Y] (<a href='/en-us/products/primary-antibodies/traf6-antibody-ep591y-ab33915'>ab33915</a>) at 1/2000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

TRAF6 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human TRAF6 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-traf6-knockout-hela-cell-line-ab266009'>ab266009</a>)

Lane 3:

HAP1 cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Predicted band size: 60 kDa

Observed band size: 65 kDa

false

Sanger Sequencing - Human TRAF6 knockout HeLa cell lysate (AB257760)
  • Sanger seq

Unknown

Sanger Sequencing - Human TRAF6 knockout HeLa cell lysate (AB257760)

Homozygous : 1 bp insertion in exon2

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2.

Disease

Adenocarcinoma

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Complementary Products

Primary Antibodies

AB33915

Anti-TRAF6 antibody [EP591Y]

RabMAb|Recombinant|KO Validated

Western blot - Anti-TRAF6 antibody [EP591Y] (AB33915)

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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

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What's included?

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Properties and storage information

Gene name
TRAF6
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

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

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.

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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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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