Anti tbk1 antibody

Manufactured by Cell Signaling Technology

Sourced in United States

The Anti-TBK1 antibody is a laboratory reagent designed for the detection and analysis of TBK1 (TANK-binding kinase 1) protein in various biological samples. TBK1 is a serine/threonine-protein kinase involved in various cellular processes, including the regulation of immune responses and inflammation. This antibody can be used in techniques such as Western blotting, immunoprecipitation, and immunohistochemistry to study the expression, localization, and interactions of TBK1 in research applications.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

Anti-TBK1 (51 citations) Anti-TBK1/NAK (E9H5S) antibody (2 citations)

5 protocols using anti tbk1 antibody

Immunoprecipitation and Immunoblotting of Phospho-TBK1

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were lysed with RIPA buffer [50 mM Tris–HCl, pH 8.0, 150 mM NaCl, 0.1% (v/v) sodium dodecyl sulfate, 1% (v/v) Nonidet-P40, and 0.04% (v/v) sodium deoxycholate] and immunoprecipitated with an anti-TBK1 antibody (Cell Signaling Technology, Danvers, MA). Precipitates were blotted using an antiphospho-TBK1 antibody (Cell Signaling Technology). After washing, primary antibodies were detected with horseradish peroxidase (HRP)-conjugated antirabbit or antimouse secondary antibodies and an ECL kit (GE Healthcare, Chicago, IL). Images of uncropped blots are provided in Supplemental data 1–3.

Hiroki H., Ishii Y., Piao J., Namikawa Y., Masutani M., Honda H., Akahane K., Inukai T., Morio T, & Takagi M. (2023). Targeting Poly(ADP)ribose polymerase in BCR/ABL1-positive cells. Scientific Reports, 13, 7588.

+ Open protocol

+ Expand

Immunoblotting of DNA Damage Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were lyzed with NETN lysis buffer (150 mM NaCl, 1 mM EDTA, 10 mM Tris-HCl, pH 8.0, 0.5% NP-40, 1 mM Na₃VO₄, 10 mM NaF, and protease inhibitor cocktail), followed by sonication for 15 seconds. After centrifugation, supernatant was resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to nitrocellulose membrane. The following antibodies were used: anti-ZMYND8 antibody (A302–089A, Bethyl Laboratories), anti-cGAS antibody (31659S, Cell Signaling Technology), anti-STING antibody (13647S, Cell Signaling Technology), anti-phospho-TBK1 antibody (5483S, Cell Signaling Technology), anti-TBK1 antibody (3013S, Cell Signaling Technology), anti-phospho-IRF3 (Ser396) antibody (29047S, Cell Signaling Technology), anti-IRF3 antibody (4302S, Cell Signaling Technology), anti-phospho-p65 antibody (3033S, Cell Signaling Technology), anti-p65 antibody (8242S, Cell Signaling Technology), anti-phospho-Chk1 (Ser296) antibody (2349S, Cell Signaling Technology), anti-Chk1 antibody (sc-8408, Santa Cruz Biotechnology), anti-γH2A.X antibody (05–636, Sigma), anti-actin antibody (A2066, Sigma), and anti-H2A.X antibody (10856–1-AP, Proteintech). Proteins were visualized by chemiluminescence with ECL prime (GE Healthcare).

Wang Y., Luo M., Chen Y., Wang Y., Zhang B., Ren Z., Bao L., Wang Y., Wang J.E., Fu Y.X., Luo W, & Wang Y. (2020). ZMYND8 expression in breast cancer cells blocks T-lymphocyte surveillance to promote tumor growth. Cancer research, 81(1), 174-186.

+ Open protocol

+ Expand

Comprehensive Antibody Optimization Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

All antibodies were used at a 1:1,000 dilution in TBS with 0.1% Tween 20 detergent buffer with 5% non-fat milk for Western blotting. Anti-Myc-Tag antibody (2278), anti-HA antibody (3724), anti-Phospho-Akt Substrate (RXRXXpS*/pT*) antibody (10001), anti-p-AKT (Ser473; 4060), anti- anti-p-p70 S6 Kinase (Thr389; 9234), anti-p-S6 Ribosomal Protein (Ser240/244; 5364), anti-p-4EBP1 (Thr37/46; 2855), pIRF3 antibody (Ser386; 37829), anti-IRF3 antibody (4302), anti-IRF7 antibody (4920), anti- anti-p-TBK1 (Ser172; 5483), anti-TBK1 antibody (51872), anti-STING antibody (13647), anti-c-Myc antibody (18583), anti-RSK1/2/3 antibody (9347), anti-Rictor antibody (9476), anti-Rbx1 (11922), anti-Skp1 antibody (12248), anti-rabbit IgG, HRP-linked antibody (7074), and anti-mouse IgG, HRP-linked antibody (7076) were obtained from Cell Signaling Technology. Anti-cyclin E antibody (sc-198), anti-β-catenin antibody (sc-59737), anti-c-Jun antibody (sc-45), anti-GST antibody (sc-459), anti-Cul1 (sc-11384), and anti-vinculin antibody (sc-25336) were obtained from Santa Cruz Biotechnology. Polyclonal anti-Flag antibody (F-7425), monoclonal anti-Flag antibody (F-3165, clone M2), and anti-α-tubulin antibody (T-5168) were obtained from Sigma-Aldrich. Anti-BUD13 antibody (20163-1-AP) and anti-Fbw7 antibody (28424-1-AP) were obtained from Proteintech.

Chen J., Zhang X., Tan X, & Liu P. (2023). Somatic gain-of-function mutations in BUD13 promote oncogenesis by disrupting Fbw7 function. The Journal of Experimental Medicine, 220(10), e20222056.

+ Open protocol

+ Expand

STING Pathway Signaling Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following antibodies were used: anti-STING antibody (Cell Signaling Technology, #50494), anti-STING pS365 antibody (Cell Signaling Technology, #72971), anti-IRF3 antibody (Cell Signaling Technology, #11904), anti-IRF3 pS396 antibody (Cell Signaling Technology, #4947), anti-TBK1 antibody (Cell Signaling Technology, #38066), anti-TBK1 pS172 antibody (Cell Signaling Technology, #5483), anti-LC3 antibody (Cell Signaling Technology, #4108), anti-p62 antibody (Cell Signaling Technology, #7695), anti-NDP52 antibody (Abcam, ab68588), anti-VAMP8 antibody (Cell Signaling Technology, #13060), anti-α-tubulin antibody (Sigma, T9026, clone DM1A), anti-FLAG antibody (Sigma, F7425 and F1804), anti-His antibody (Cell Signaling Technology, #2366), and anti-GFP antibody (Proteintech, 50430-2-AP). All secondary antibodies were purchased from Jackson ImmunoResearch.

Song X., Xi Y., Dai M., Li T., Du S., Zhu Y., Li M., Li Y., Liu S., Ding X., Yao X., Lai Y, & Liu X. (2024). STING guides the STX17-SNAP29-VAMP8 complex assembly to control autophagy. Cell Insight, 3(2), 100147.

+ Open protocol

+ Expand

Protein Extraction and Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were lysed and total protein was followingly extracted by RIPA lysis buffer (Beyotime), with the concentration evaluated by a BCA protein assay kit (Beyotime). Protein samples were followingly mixed with loading buffer, denatured, and separated by SDS‐PAGE and transferred onto PVDF membranes (Millipore, Bedford, MA, USA), which was blocked with 5% skimmed milk for 1 h at ambient temperature and incubated with the corresponding primary antibodies: anti‐Iba‐1 antibody (17198; Cell Signaling Technology, MA, USA), anti‐β‐actin antibody (8457; Cell Signaling Technology), anti‐STING antibody (13647; Cell Signaling Technology), anti‐phospho‐TANK binding kinase 1 (TBK1) antibody (5483; Cell Signaling Technology), and anti‐TBK1 antibody (3504; Cell Signaling Technology) overnight at 4°C, followed by incubation with HRP‐conjugated secondary antibody (Beyotime) for 1.5 h at ambient temperature. The protein bands were developed with an enhanced chemiluminescence kit (Beyotime). Ultimately, the relative expressions of proteins were analyzed by Image J software (NIH, USA) with β‐actin as the internal reference.

Song N., Song R, & Ma P. (2022). MiR‐340‐5p alleviates neuroinflammation and neuronal injury via suppressing STING in subarachnoid hemorrhage. Brain and Behavior, 12(9), e2687.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!