TRAF2 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: 7 bp deletion in exon 2.
E3 ubiquitin-protein ligase TRAF2, MGC:45012, OTTHUMP00000022625, OTTHUMP00000064745, TNF receptor-associated factor 2, TNF receptor-associated protein, TR AP, TRAF2_HUMAN, TRAP 3, Tumor necrosis factor type 2 receptor-associated protein 3
TRAF2 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: 7 bp deletion in exon 2.
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.
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.
TRAF2 known also as TNF Receptor-Associated Factor 2 is an adapter protein weighing approximately 56 kDa. It is expressed in many tissues such as lymph nodes spleen and thymus. This protein acts as a signal transducer playing an integral role in the activation of NF-kB and MAPK signaling pathways. TRAF2 interacts with TNF receptors and other proteins using its RING finger and zinc finger domains for ubiquitination processes.
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.
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.
TRAF2’s dysregulation is linked with autoimmune diseases and certain types of cancer. Overactivity can contribute to chronic inflammation highlighting its role in autoimmune disorders like rheumatoid arthritis. In cancer TRAF2’s involvement with TNF receptors can lead to uncontrolled cell proliferation. Proteins like cIAP1 and cIAP2 are often related to TRAF2 in these contexts influencing the progression and modulation of disease states.
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Anti-TRAF2 antibody [EPR6048] 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 Human TRAF2 knockout HEK-293T 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. Anti-TRAF2 antibody [EPR6048] ab126758 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (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] (Anti-TRAF2 antibody [EPR6048] ab126758) 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)
Performed under reducing conditions.
Predicted band size: 55 kDa
Observed band size: 55 kDa
Anti-TRAF2 antibody [EPR7064] 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 Human TRAF2 knockout HEK-293T 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. Anti-TRAF2 antibody [EPR7064] ab167163 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (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] (Anti-TRAF2 antibody [EPR7064] ab167163) 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)
Performed under reducing conditions.
Predicted band size: 55 kDa
Observed band size: 55 kDa
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.
Homozygous: 7 bp deletion in exon 2
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