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Ab40676

Manufactured by Abcam
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Sourced in United Kingdom

Ab40676 is a primary antibody product offered by Abcam. It is a polyclonal antibody that recognizes a specific target. The core function of this product is to detect and bind to the target antigen for research purposes.

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Lab products found in correlation

Ab109272, by Abcam (5 mentions) Lipopolysaccharides (lps), by Merck Group (3 mentions) Hek293 cell, by American Type Culture Collection (2 mentions) Ab52915, by Abcam (2 mentions) Ab8245, by Abcam (2 mentions) Ab183218, by Abcam (2 mentions) Ab168352, by Abcam (2 mentions) Cd163, by BioLegend (2 mentions) Phospho nf κb, by Cell Signaling Technology (2 mentions) Ripk3, by Cell Signaling Technology (2 mentions) Ab178945, by Wuhan Servicebio Technology (2 mentions) Hrp conjugated anti rabbit and anti mouse secondary antibodies, by Boster Bio (2 mentions) Nf κb, by Proteintech (2 mentions) Phospho tbk1 nak ser172, by Cell Signaling Technology (2 mentions) Gapdh, by Wuhan Servicebio Technology (2 mentions) M csf, by BioLegend (2 mentions) Phospho ripk3, by Cell Signaling Technology (2 mentions) F4 80, by BioLegend (2 mentions) Bx53 microscope, by Olympus (1 mentions) Ab93159, by Abcam (1 mentions)

16 protocols using ab40676

1

Immunohistochemical Analysis of TBK1 in Human Retina

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Human retina samples were collected in the Tongji Hospital. The present study was approved by the Research Ethics Committee of the Tongji Hospital, Huazhong University of Science and Technology, and individual permission was obtained using standard informed consent procedures. The investigation conformed to the principles outlined in the Declaration of Helsinki regarding the use of human tissues. Preparing for immunohistochemistry, samples were fixed with 4% PFA, embedded in paraffin, and sectioned at 4 μm. After treatment with 3% H2O2 to eliminate endogenic peroxidase, the sections were incubated with an anti-TBK1 antibody (Abcam, ab40676). Thereafter, the sections were stained with horseradish peroxidase-conjugated secondary antibodies for 1 h. Images were captured using an Olympus BX53 microscope.
Zhao B., Ni Y., Zhang H., Zhao Y, & Li L. (2022). Endothelial deletion of TBK1 contributes to BRB dysfunction via CXCR4 phosphorylation suppression. Cell Death Discovery, 8, 429.
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2

Antibody Characterization and Signaling Pathway Analysis

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Mouse monoclonal antibodies against Flag (Origene, 1:2000, F3165), HA (Origene, 1:2000, H6908), β-actin (Sigma, 1:10,000, A2228), Myc (CST, 1:1000, 5605), p-IκBα (CST, 1:1000, 9246 L); rabbit antibodies against p-IRF3 (CST, 1:500, 37829), USP19 (Abcam, 1:1000, ab93159), TRIF (Abcam, 1:1000, ab180689), p65 (Santa Cruz Biotechnology, 1:1000, 71675), p-p65(S536) (CST, 1:1000, 3033),TBK1 (Abcam, 1:1000, ab40676) and p-TBK1 (Abcam, 1:1000, ab109272), ubiquitin (Abcam, 1:500, ab134953), K27-linkage specific polyubiquitin (Abcam, 1:1000, 181537), TLR3 (CST, 1:500, 6961), TLR4 (R&D, 1:500, AF1478), TRAM (Abcam, 1:500, ab96106), KCTD10 (Proteintech, 1:1000, 27279–1-AP); poly(I:C) (Invivogen), LPS (Sigma), R848 (Invivogen), PGN (Invivogen), human IFN-γ (Peprotech), murine M-CSF (Peprotech), Trizol (Takara Bio), SYBR Green (BIO-RAD), dual-specific luciferase assay kit (Promega, E1980), polybrene (Millipore,TR-1003-G), type II collagenase (Worthington), DNase I (Sigma-Aldrich), and d-galactosamine hydrochloride (D-Gal) (Sigma); and ELISA kits for TNF (Biolegend), IL-6 (Biolegend), CXCL10 (Boster) and IFN-β (PBL) were purchased from the indicated companies. HEK293 cells were obtained from ATCC. SeV (Cantell strain) (Charles River Laboratories), HSV-1 (KOS strain) (China Center for Type Culture Collection, Wuhan, China) were obtained from the indicated companies.
Wu X., Lei C., Xia T., Zhong X., Yang Q, & Shu H.B. (2019). Regulation of TRIF-mediated innate immune response by K27-linked polyubiquitination and deubiquitination. Nature Communications, 10, 4115.
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3

Comprehensive Reagents for Cellular Signaling Studies

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CTB-Alexa 555 was bought from BrainVTA (Wuhan, Hubei, China). Degrasyn and Amlexanox were purchased from MedChemExpress (Monmouth Junction, NJ, USA). Proteasome inhibitor MG132 (Calbiochem), cycloheximide (CHX; Sigma‐Aldrich). Antibodies: anti-NAK/TBK1 (ab40676; Abcam); anti-NAK/TBK1 (ab109272; Abcam); anti-USP9X (ab180191; Abcam); anti-synaptophysin (ab14692; Abcam); anti-p70S6k (2708T; Cell Signaling Technology); anti-p-p70S6k (9234T; Cell Signaling Technology); anti-GAPDH (10494-1-AP; Proteintech);anti-HA (TT0008; Abmart); anti-GFP (M20004; Abmart); anti-MYC (M20002; Abmart); anti-His (M20001; Abmart); anti-Flag (TT0003; Abmart); anti-Flag (66008-4-Ig; Proteintech); anti-β-actin (sc-47778; Santa Cruz); anti-RAPTOR (20984-1-AP; Proteintech); anti-Tuj1 (801201; Biolegend); anti-Iba1 (ab48004; Abcam); anti-GAFP (ab4674; Abcam); and anti-pS6(Ser240/244) (5364T; Cell Signaling Technology).
Ye M., Hu Y., Zhao B., Mou Q., Ni Y., Luo J., Li L., Zhang H, & Zhao Y. (2023). TBK1 Knockdown Alleviates Axonal Transport Deficits in Retinal Ganglion Cells Via mTORC1 Activation in a Retinal Damage Mouse Model. Investigative Ophthalmology & Visual Science, 64(10), 1.
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4

Western Blot Analysis of Macrophage Proteins

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The proteins were isolated from liver macrophages and cultured RAW264.7 cells by lysing them in RIPA buffer containing 1% PMSF (Beyotime). The protein concentration was determined using a BCA protein kit (Beyotime). Based on the protein concentration, equal amounts of protein were separated by SDS/PAGE and transferred onto a PVDF membrane (Millipore Corp.) followed by blocking. The following primary antibodies were used in this study: anti‐STING (19851‐1‐AP; Proteintech), anti‐BAX (ab32503; Abcam), anti‐BAK (#578; CST), anti‐Bcl2 (ab182858; Abcam), anti‐p65 (#8242S; CST), anti‐p‐p65 (#3033S: CST), anti‐TBK1 (ab40676; Abcam), anti‐p‐TBK1 (#5483S; CST), anti‐caspase3 (ab184787; Abcam), anti‐IRF3 (#11904S; CST), anti‐p‐IRF3 (#37829S; CST), and anti‐β‐actin (7D2C10; Proteintech). The protein bands were visuzed using an enhanced chemiluminescence detection system (Bio‐Rad) and quantified after normzation to internal control β‐actin using the ImageJ software (NIH).
Wang L., You H., Meng H., Pan X., Chen X., Bi Y., Zhang Y., Li J., Yin N., Zhang Z., Huang C, & Li J. (2021). STING‐mediated inflammation contributes to Gao binge ethanol feeding model. Journal of Cellular Physiology, 237(2), 1471-1485.
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5

Immunocytochemical and Western Blot Analyses

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For immunocytochemical studies, the following primary antibodies were used: LC3 mouse monoclonal (M152-3/1:50; MBL), Iba1 rabbit polyclonal (019-19741/1:1,000; Wako), and Lamp1 rat polyclonal (1D4B/1:300; DHSB). Alexa Fluor secondary antibodies were from ThermoFisher (1:400). For studies involving Western blots, the following primary antibodies were used: LC3 mouse monoclonal (M186-3/1:1,000; MBL), actin (A2228/1:5,000; Sigma), p-AMPK (T172) rabbit polyclonal (2535/1:1,000; CST), AMPK rabbit polyclonal (5831/1:1,000; CST), p-TBK1 (S172) rabbit polyclonal (5483/1:1,000; CST), TBK1 rabbit polyclonal (ab40676/1:1,000; Abcam), p-p62 (S403) rat polyclonal (D343-3/1:1,000; MBL), p62 mouse monoclonal (ab56416/1:1,000; Abcam), p-S6K (T389) rabbit polyclonal (AT-7159/1:1,000; MBL), S6K rabbit polyclonal (2708/1:1,000; CST), GFP chicken polyclonal (ab13970/1:5,000; Abcam), and mCherry rabbit polyclonal (ab167453/1:1,000; Abcam). For FACS, the following antibodies were used: Alexa Flour 488–conjugated anti-NeuN (MAB377X/1:1,000; Millipore), PE-conjugated anti-GFAP (561483/1:50; BD Biosciences), and APC-conjugated anti-Cd11b (561690/1:250; BD Biosciences).
Chang M.C., Srinivasan K., Friedman B.A., Suto E., Modrusan Z., Lee W.P., Kaminker J.S., Hansen D.V, & Sheng M. (2017). Progranulin deficiency causes impairment of autophagy and TDP-43 accumulation. The Journal of Experimental Medicine, 214(9), 2611-2628.
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6

Antibody Characterization for TBK1 and IRF3

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The antibody against TBK1 (ab40676) was purchased from Abcam. Antibodies against cGAS (#31659), IRF3 (#4302), phospho-IRF3 (#4947), and phospho-TBK1 (#5483) were purchased from Cell Signaling Technology. The anti-GAPDH (sc-32233) antibody was purchased from Santa Cruz Biotechnology. Anti-MB21D1 (HPA031700) and anti-β-actin (A5441) antibodies were purchased from Sigma-Aldrich. The cyanine 3-conjugated secondary antibody (goat anti-rabbit) was purchased from Jackson ImmunoResearch.
Herring testis DNA (HT-DNA) was purchased from Sigma-Aldrich. HSV-1 and HSV-1-GFP were kindly provided by Dr. Wentao Qiao (Nankai University) and Dr. Chunfu Zheng (Suzhou University), respectively. HSV-1 was propagated and titered by plaque assays in Vero cells.
Yu Q., Chu L., Li Y., Wang Q., Zhu J., Wang C, & Cui S. (2021). miR-23a/b suppress cGAS-mediated innate and autoimmunity. Cellular and Molecular Immunology, 18(5), 1235-1248.
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7

Progesterone and Dasatinib Modulate IFN Signaling

Cited 1 time
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Progesterone (HY-N0437) and Dasatinib (HY-10181) (MedChemExpress); IFN-γ (300-02, Peptech); Anti-mouse IFNAR-1-in vivo (clone MAR1-5A3; Selleck A2121); ELISA kits for progesterone (582601, Cayman Chemical), GnRH (MM-0506M1) and LH (MM-44039M1) (MEIMAIN), murine IFN-β(42400, PBL), murine CXCL10 (EMC121, Neobioscience), murine TNFα (430904, BioLegend), murine IL-6 (431304, BioLegend); Mouse monoclonal antibodies against HA (TA180128, OriGene), Flag (F3165) and β-actin (A2228) (Sigma), IRF3 (sc-33641, Santa Cruz Biotechnology); Rabbit polyclonal antibodies against PGR (8757), SRC (2109), p-SRCY419 (2101) and p-Tyrosine (9411S) (Cell signaling technology), TBK1 (ab40676), p-TBK1S172 (ab109272) and p-IRF3S386 (ab76493) (Abcam) were purchased from the indicated companies. SeV, EMCV and HSV-1 were previously described.75 (link),76 (link) HEK293 cells were obtained from ATCC. The T-47D cells were provided by Dr. Jing Zhang (Wuhan University). The Calu-3 cells were provided by Dr. Ke Lan (Wuhan University).
Su S., Hua D., Li J.P., Zhang X.N., Bai L., Cao L.B., Guo Y., Zhang M., Dong J.Z., Liang X.W., Lan K., Hu M.M, & Shu H.B. (2022). Modulation of innate immune response to viruses including SARS-CoV-2 by progesterone. Signal Transduction and Targeted Therapy, 7, 137.
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8

Antibody-Based Immunoblotting Protocol

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The antibodies used for immunoblotting include: mouse monoclonal antibodies (mAbs) c-Myc antibody (sc-56634) and β-actin (C4) (sc-47778) (both purchased from Santa Cruz Biotechnology, Dallas, TX, USA); rabbit polyclonal antibodies against NP (GTX125989) was purchased from GeneTex (Taiwan); anti-TBK1 (ab40676), and anti-TBK1 (phosphor S172) (ab109272) antibodies were obtained from Abcam (Cambridge, MA, USA). Rabbit polyclonal antibody against acetylated-lysine (9441), mouse mAb against HDAC1 (10E2) (5356), p-IRF3 (4947), IRF3 (4302), p-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (4370), p44/42 MAPK (Erk1/2) (9102), and rabbit mAb HA-Tag (c29F4) (28) were from Cell Signaling Technology (Beverly, MA, USA).
Chen L., Wang C., Luo J., Su W., Li M., Zhao N., Lyu W., Attaran H., He Y., Ding H, & He H. (2017). Histone Deacetylase 1 Plays an Acetylation-Independent Role in Influenza A Virus Replication. Frontiers in Immunology, 8, 1757.
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9

Characterization of cGAS-STING Pathway

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Western blotting was performed as previously described [42 (link)]. Briefly, cells were lysed in cold radio immunoprecipitation assay (RIPA) lysis buffer for 30 min and centrifuged to remove precipitates. Loading buffer was added, then samples were boiled for 10 min. Proteins were separated by SDS-PAGE and transferred to PVDF membranes, which were washed with Tris-buffered saline plus Tween-20 (TBST) and blocked with 5% non-fat milk. Next, the membranes were incubated with primary antibodies overnight at 4 °C, washed with TBST, then incubated with horseradish peroxidase (HRP)-labeled secondary antibodies. Finally, an enhanced chemiluminescence system was used to detect proteins according to the manufacturer’s protocol. The antibodies used were rabbit anti-cGAS (D3080, CST, anti-MB21D1, Boston, MA, USA), rabbit anti-STING (19851-1-AP, Proteintech, Chicago, IL, USA), rabbit anti-TBK1 (ab40676, Abcam, Cambridge, UK), and p-TBK1 (ab109272, Abcam).
Li Q., Liu C., Yue R., El-Ashram S., Wang J., He X., Zhao D., Zhou X, & Xu L. (2019). cGAS/STING/TBK1/IRF3 Signaling Pathway Activates BMDCs Maturation Following Mycobacterium bovis Infection. International Journal of Molecular Sciences, 20(4), 895.
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10

Comprehensive Immune Signaling Pathway Analysis

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Anti-KI67 (abcam, ab16667), Anti-KI67 (ab279653), Anti-TOMM20 (abcam, ab186735), Anti-FTO (ab126605), Anti-Vimentin (Santa, sc-80975), Anti-COL II (abcam, ab34712), Anti-MMP3 (abcam, ab52915), Anti-MMP13 (abcam, ab39012), Anti-dsDNA (abcam, ab27156), Anti-CMPK2 (Proteintech, 25877-1-AP), Anti-STING (CST.13647), Anti-cGAS (Proteintech, 29958-1-AP), Anti-TBK1 (abcam, ab40676), Anti-p-TBK1 (CST.5483), Anti-p-IRF3 (CST.29057), Anti-IL-1β (abcam, ab315084), Anti-IL-6 (abcam, ab290735), Anti-GADPH (abcam, ab8245).
Jin L., Chen Q., Hu K., Fan D., Zhang H., Deng J., Qi W, & Yu Q. (2024). The FTO-CMPK2 Pathway in Fibroblast-like Synoviocytes Modulates Rheumatoid Arthritis Synovial Inflammation and Cartilage Homeostasis via mtDNA Regulation. International Journal of Biological Sciences, 20(5), 1617-1633.
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