Testing Data
(Fig-1: Induction of RIG-I activity by poly(I: C) prepacked with lipofectamine in RIG-I Leeporter HEK293T cells.)
RIG-I/NF-kB cell line
RIG-I/NF-kB Leeporter HEK293T Cell Line
Testing Data
(Fig-1: Induction of RIG-I activity by poly(I: C) prepacked with lipofectamine in RIG-I Leeporter HEK293T cells.)
Testing Data
(Fig-1: Induction of RIG-I activity by poly(I: C) prepacked with lipofectamine in RIG-I Leeporter HEK293T cells.)
NCBI and Uniprot Product Information
Uniprot Description
RIPK1: Serine-threonine kinase which transduces inflammatory and cell-death signals (necroptosis) following death receptors ligation, activation of pathogen recognition receptors (PRRs), and DNA damage. Upon activation of TNFR1 by the TNF-alpha family cytokines, TRADD and TRAF2 are recruited to the receptor. Ubiquitination by TRAF2 via 'Lys-63'-link chains acts as a critical enhancer of communication with downstream signal transducers in the mitogen-activated protein kinase pathway and the NF-kappa-B pathway, which in turn mediate downstream events including the activation of genes encoding inflammatory molecules. Polyubiquitinated protein binds to IKBKG/NEMO, the regulatory subunit of the IKK complex, a critical event for NF-kappa-B activation. Interaction with other cellular RHIM-containing adapters initiates gene activation and cell death. RIPK1 and RIPK3 association, in particular, forms a necroptosis-inducing complex. Interacts (via RIP homotypic interaction motif) with RIPK3 (via RIP homotypic interaction motif); this interaction induces RIPK1 necroptosis-specific phosphorylation, formation of the necroptosis-inducing complex. Interacts (via the death domain) with TNFRSF6 (via the death domain) and TRADD (via the death domain). Is recruited by TRADD to TNFRSF1A in a TNF-dependent process. Binds RNF216, EGFR, IKBKG, TRAF1, TRAF2 and TRAF3. Interacts with BNLF1. Interacts with SQSTM1 upon TNF-alpha stimulation. May interact with MAVS/IPS1. Interacts with ZFAND5. Interacts with RBCK1. Interacts with BIRC2/c-IAP1, BIRC3/c-IAP2 and XIAP/BIRC4. Inhibited by necrostatin-1. Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family. 2 isoforms of the human protein are produced by alternative splicing.
Protein type: EC 2.7.11.1; Kinase, protein; Protein kinase, Ser/Thr (non-receptor); Protein kinase, TKL; RIPK family; TKL group
Chromosomal Location of Human Ortholog: 6p25.2
Cellular Component: cytoplasm; cytosol; mitochondrion; receptor complex
Molecular Function: death receptor binding; identical protein binding; protein binding; protein complex binding; protein kinase activity; protein serine/threonine kinase activity; signal transducer activity; ubiquitin protein ligase binding
Biological Process: activation of JNK activity; activation of NF-kappaB transcription factor; apoptosis; caspase activation; cellular protein catabolic process; I-kappaB kinase/NF-kappaB cascade; negative regulation of I-kappaB kinase/NF-kappaB cascade; positive regulation of apoptosis; positive regulation of I-kappaB kinase/NF-kappaB cascade; positive regulation of interleukin-8 production; positive regulation of JNK cascade; positive regulation of macrophage differentiation; positive regulation of programmed cell death; positive regulation of protein amino acid phosphorylation; positive regulation of transcription from RNA polymerase II promoter; positive regulation of tumor necrosis factor production; protein amino acid autophosphorylation; protein heterooligomerization; protein homooligomerization; tumor necrosis factor-mediated signaling pathway
Research Articles on RIG-I/NF-kB
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Product Notes
The RIG-I/NF-kB ripk1 (Catalog #AAA668873) is a Cell Line and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's RIG-I/NF-kB can be used in a range of immunoassay formats including, but not limited to, Functional Assay. Monitor the RIG-I signaling pathway activity. Screen for activators or inhibitors of the RIG-I signaling pathway. Culture conditions: Cells should be grown at 37 degree C with 5% CO2 using DMEM medium supplemented with 10% FBS and 1% Pen/Strep, plus 2 ug/ml Puromycin and 5 ug/ml blasticidin. It is recommended to quickly thaw the frozen cells upon receipt or from liquid nitrogen in a 37 degree C water-bath, transfer to a tube containing 10 ml of growth medium without Puromycin and Blasticidin, spin down cells, resuspend cells in pre-warmed growth medium without Puromycin and Blasticidin, transfer resuspended cells to T25 flask and culture in 37 degree C-CO2 incubator. Leave the T25 flask in the incubator for 2~4 days without disturbing or changing the medium until cells completely recover viability and become adherent. Once cells are over 90% adherent, remove growth medium and passage the cells through trypsinization and centrifugation. At first passage, switch to growth medium containing Puromycin and Blasticidin. Cells should be split before they reach complete confluence. To passage the cells, detach cells from culture vessel with Trypsin/EDTA, add complete growth medium and transfer to a tube, spin down cells, resuspend cells and seed appropriate aliquots of cells suspension into new culture vessels. Subcultivation ration = 1: 10 to 1: 20 weekly. Functional validation: A. Response of RIG-I Leeporter- HEK293T cells to Poly(I: C). 1. Harvest RIG-I Leeporter- HEK293T cells and seed cells into a white solid-bottom 96-well microplate in 100 ul of growth medium at 5 x 10^4 cells/well. 2. Incubate cells at 37 degree C in a CO2 incubator for overnight. 3. The next day, stimulate cells with various concentrations of the poly(I: C) packed with lipofectamine 2000, which was prepared as follows: i) A 50 ul of 10 mg/ml poly(I: C) (= 500 ug total) is preincubated in 50 ul Opti-MEM (Life Technologies) for 5 min. ii) Similarly, a 20 ul Lipofectamine 2000 (Life Technologies) is preincubated in 80 ul Opti-MEM for 5 min. iii) After 5 min, they are combined together as a total volume of 200 ul and further incubated for 20 min at room temperature. iv) The poly(I: C) packed with Lipofectamine 2000 (A 200 ul total at 2.5 mg/ml) is then used to stimulate cells. 4. Incubate at 37 degree C in a CO2 incubator for 6-16 hours. 5. Add 30-50 ul of luciferase assay reagent per well. 6. Incubate at room temperature for 1-5 minutes and measure luminescence using a microplate luminometer. Researchers should empirically determine the suitability of the RIG-I/NF-kB ripk1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "RIG-I/NF-kB, Cell Line" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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