Human TRAF6 knockout HeLa cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human TRAF6 knockout HeLa cell line (AB266009)
Lanes 1- 4 : Merged signal (red and green). Green - ab40675 observed at 65 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab40675 was shown to 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 HeLa and TRAF6 knockout HeLa cell lysates 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) overnight at 4°C at a 1 in 500 dilution and a 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 (ab266009)
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
- WB
Lab
Western blot - Human TRAF6 knockout HeLa cell line (AB266009)
Lanes 1- 4 : Merged signal (red and green). Green - ab33915 observed at 65 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab33915 was shown to 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 HeLa and TRAF6 knockout HeLa cell lysates 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) overnight at 4°C at a 1 in 2000 dilution and a 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 (ab266009)
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 seq
Unknown
Sanger Sequencing - Human TRAF6 knockout HeLa cell line (AB266009)
Homozygous : 1 bp insertion in exon 2.
Complementary Products
Primary Antibodies
AB33915
RabMAb|Recombinant|KO Validated|20ul selling size
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Primary Antibodies
AB138491
Anti-GST3 / GST pi antibody [EPR8263]
RabMAb|Recombinant|KO Validated
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Reactivity data
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?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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.
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.
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 promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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