Human TRAF2 knockout HEK-293T cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human TRAF2 knockout HEK-293T cell line (AB266060)
Lanes 1- 2 : Merged signal (red and green). Green - ab167163 observed at 55 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab167163 was shown to react with TRAF2 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266060 (knockout cell lysate ab257759) was used. Wild-type HEK-293T and TRAF2 knockout HEK-293T 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. ab167163 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4° at a 1 in 1000 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-TRAF2 antibody [EPR7064] (<a href='/en-us/products/primary-antibodies/traf2-antibody-epr7064-ab167163'>ab167163</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
TRAF2 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human TRAF2 knockout HEK-293T cell line (ab266060)
Predicted band size: 55 kDa
Observed band size: 55 kDa
false
- WB
Lab
Western blot - Human TRAF2 knockout HEK-293T cell line (AB266060)
Lanes 1- 2 : Merged signal (red and green). Green - ab126758 observed at 55 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab126758 was shown to react with TRAF2 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266060 (knockout cell lysate ab257759) was used. Wild-type HEK-293T and TRAF2 knockout HEK-293T 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. ab126758 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4° at a 1 in 1000 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-TRAF2 antibody [EPR6048] (<a href='/en-us/products/primary-antibodies/traf2-antibody-epr6048-ab126758'>ab126758</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
TRAF2 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human TRAF2 knockout HEK-293T cell line (ab266060)
Predicted band size: 55 kDa
Observed band size: 55 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human TRAF2 knockout HEK-293T cell line (AB266060)
Homozygous : 7 bp deletion in exon 2
- Cell Culture
Lab
Cell Culture - Human TRAF2 knockout HEK-293T cell line (AB266060)
Representative images TRAF2 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.
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![Flow Cytometry (Intracellular) - Anti-TRAF2 antibody [EPR6048] (AB126758)](https://content.abcam.com/products/images/traf2-antibody-epr6048-ab126758--flow-cytometry-intracellular-img75285.jpg)
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 and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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
TRAF2 modulates immune responses by regulating cell survival proliferation and apoptosis. It often forms complexes with other TRAF family proteins to mediate signal transmission. These complexes control the balance between pro-apoptotic and anti-apoptotic signals impacting the regulation of immune and inflammatory responses. Its interactions are necessary for the proper function of NF-kB ensuring balanced immune activities.
Pathways
TRAF2 is a critical component within both NF-kB and JNK signaling pathways. In the NF-kB pathway TRAF2 intersects with proteins like IKK and NEMO facilitating the transcription of genes involved in immune and inflammatory responses. In JNK signaling TRAF2 influences cellular responses through regulation of apoptotic signals. By linking these pathways TRAF2 ensures a coordinated cellular response to extracellular stimuli.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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