TBK1 Overexpression Lysate Summary
Description |
TBK1 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC205238
Accession#: NM_013254
Protein Tag: C-MYC/DDK
You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT. |
Gene |
TBK1 |
Applications/Dilutions
Dilutions |
|
Application Notes |
This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag ( NBP1-71705) present on the protein construct. Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading. |
Theoretical MW |
83.5 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. |
Packaging, Storage & Formulations
Storage |
Store at -80C. Avoid freeze-thaw cycles. |
Buffer |
RIPA buffer |
Lysate Details for Array
Notes
HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.
Alternate Names for TBK1 Overexpression Lysate
Background
NFkappaB-activating kinase (NAK) also known as TANK binding kinase-1 (TBK1) or tumor necrosis factor receptor associated factor 2-associated kinase (T2K) is a serine/threonine protein that belongs to the IkB kinase (IKK) family (1). Similar to IKK alpha and beta (2), NAK induces IkB degradation and NF-kB activity. NAK is activated by phorbol ester tumour promoters and growth factors, whereas catalytically inactive NAK specifically inhibits activation of NF-kB by protein kinase C-epsilon (PKCepsilon). NAK specifically phosphorylates IkB alpha on Serine 36 and NFkB subunit RelA (p65) on Serine 536 (3). NAK is also known to interact with TANK, TRAF-2 and TRIF (4).
Limitations
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are
guaranteed for 6 months from date of receipt.
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Publications for TBK1 Lysate (NBP2-10346) (0)
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Product General Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
Video Protocols
FAQs for TBK1 Lysate (NBP2-10346). (Showing 1 - 6 of 6 FAQ).
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What is the exact sequence this TBK1 antibody recognizes?
- The exact epitope of this TBK1 antibody is unknown because we do not perform epitope mapping. This TBK1 antibody was made against a synthetic peptide corresponding to aa 563-577 of human TBK1.
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I'm using NAX (TBK1 antibody) why is my band different from the predicted molecular weight?
- While this TBK1 antibody is typically observed around 83 kDA variations including PTMs, relative charges and other experimental factors can affect where the band is observed.
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We are looking to use your TBK1 antibody in simple western but need a good loading control also validated in simple western, which would you recommend?
- Our alpha tubulin loading control antibody (NB100-690) would be a good choice to use with the TBK1 antibody; it detects around 55 kDa and is validated in simple western.
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Is there a smaller size of your TBK1 antibody?
- This TBK1 antibody is offered in a smaller 0.025 mg size.
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Are there any publications where this TBK1 antibody was used in western blot for mouse?
- This TBK1 antibody was cited in a study looking at the relation between loss of TBK1 activity and increased susceptibility to LPS induced lethality. PMID 20651301
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Cant this TBK1 antibody be conjugated to PE for FLOW?
- Although it has not been validated for flow we do offer this TBK1 antibody as a PE conjugate. If you would like to test in FLOW you may want to take advantage of our innovator's reward program.