Total tbk1

Manufactured by Cell Signaling Technology

Total TBK1 is a laboratory reagent that can be used to quantify the total amount of TBK1 (TANK-binding kinase 1) protein in a sample. TBK1 is a serine/threonine protein kinase involved in various cellular signaling pathways, including the regulation of immune and inflammatory responses. The Total TBK1 product provides a reliable and standardized method for measuring the total levels of TBK1 in biological samples, such as cell lysates or tissue extracts.

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Lab products found in correlation

Total nf κb p65, by Cell Signaling Technology (1 mentions) Phospho nf κb p65, by Cell Signaling Technology (1 mentions) P tbk1, by Cell Signaling Technology (1 mentions) Sting, by Cell Signaling Technology (1 mentions) Mrt67307, by Merck Group (1 mentions) Phospho histone h3, by Cell Signaling Technology (1 mentions) Phospho plk1, by Cell Signaling Technology (1 mentions) Amlexanox, by Bio-Techne (1 mentions) Bx795, by Selleck Chemicals (1 mentions) α tubulin, by Merck Group (1 mentions) Nms p715, by Merck Group (1 mentions) Control sirna sc 37007, by Santa Cruz Biotechnology (1 mentions) Sc 13162, by Santa Cruz Biotechnology (1 mentions)

2 protocols using total tbk1

Western Blot Analysis of Immune Signaling Proteins

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Cells were washed and lysed with buffer containing 50 mM Tris-HCl (pH 7.6), 150 mM NaCl, 0.1% SDS, 1% NP-40, and protease/phosphatase inhibitors. Protein concentration was determined by Bradford Pierce BCA protein assay (Thermo Fisher Scientific) according to the manufacturer’s instructions. The supernatants were boiled for 10 minutes and separated by SDS-PAGE. Polyacrylamide gels were cast with GERBU acrylamide M-Bis 30% solution; depending on size, proteins were spread on running gel from 10% to 12%.
Primary antibodies included WASp (Santa Cruz Biotechnology Inc., B-9), phospho–NF-κB p65 (Cell Signaling Technology, 93H1), total NF-κB p65 (Cell Signaling Technology, C22B4), p-TBK1 (Cell Signaling Technology, 52C2), total TBK1 (Cell Signaling Technology, D52C2), STING (Cell Signaling Technology, D2P2F).
Secondary antibodies included anti-rabbit HRP and anti-mouse HRP (Jackson ImmunoResearch Laboratories Inc.).

Piperno G.M., Naseem A., Silvestrelli G., Amadio R., Caronni N., Cervantes-Luevano K.E., Liv N., Klumperman J., Colliva A., Ali H., Graziano F., Benaroch P., Haecker H., Hanna R.N, & Benvenuti F. (2020). Wiskott-Aldrich syndrome protein restricts cGAS/STING activation by dsDNA immune complexes. JCI Insight, 5(17), e132857.

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Inhibition of TBK1, TTK, and APC/C in Cell Studies

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Cells were treated with BX795 (#s1274, selleckchem), MRT67307 (#SML0702, Millipore Sigma) or amlexanox (#4857, Tocris) to inhibit TBK1 and NMS-P715 (#475949, Millipore Sigma) to inhibit TTK Plasmids expressing GST-Cdh1 and GST-Cdc20 were kind gift from Dr. Lixin Wan. Phospho (S172) TBK1 (#5483), phospho PLK1 (#5472S), phospho histone H3 (#3377), TTK (#3255T), APC1 (#13329), pAurora A (Thr288), B (Thr232), C (Thr 198) (#2914S) and total TBK1 (#3013S) antibodies were purchased from Cell Signaling, and antibody against Cdc20 (SC-13162) was purchased from Santa Cruz Biotechnology. β-actin (#A1978, clone AC-15) and α-tubulin (#T6074) antibodies were purchased from Sigma Chemical Co. Cyclin B1 antibody (#5472S) was purchased from BD Biosciences. pTTK antibody (#44–1325G) was purchased from Novex. BubR1 (ab70544), Mad2 (ab97777) and Cdh1 (ab89535) antibodies were purchased from Abcam. γ-tubulin antibody (# PA5–34815) was purchased from Invitrogen. Control siRNA (sc-37007) was purchased from Santa Cruz Biotechnology and TBK1 siRNA (# 4457298, ID: s761) was purchased from Ambion.

Maan M., Agrawal N.J., Padmanabhan J., Leitzinger C.C., Rivera-Rivera Y., Saavedra H.I, & Chellappan S.P. (2020). Tank Binding Kinase 1 modulates spindle assembly checkpoint components to regulate mitosis in breast and lung cancer cells. Biochimica et biophysica acta. Molecular cell research, 1868(3), 118929.

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