Anti tbc1d1

Manufactured by Cell Signaling Technology

Anti-TBC1D1 is a primary antibody product from Cell Signaling Technology. It is used to detect the TBC1D1 protein, which is involved in cellular signaling pathways.

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4 protocols using anti tbc1d1

Antibodies for Western Blot Analysis

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All primary antibodies used for western blots were purchased from commercial sources. Antibodies from Santa Cruz include: anti-beclin 1 (sc11427), anti-myc HRP (sc40 HRP) and anti-β-actin HRP (sc47778 HRP). Antibodies from Cell Signaling Technology include: anti-UVRAG (#11315), anti-Vps34 (#4263), anti-Atg14 (#96752, to detect mouse Atg14), anti-p-AMPKα(Thr172) (#2535), anti-AMPKα (#2532), anti-p-Raptor(Ser792) (#2083), anti-Raptor (#2280), anti-TBC1D1 (#66433) and anti-HA (#3724, to detect Tlr9-HA in tissue lysates of Tlr9-HA KI mice). Other primary antibodies include: anti-p-ACC(Ser79) (#07–303, Millipore), anti-ACC (#04–322, Millipore), anti-p-TBC1D1(Ser237) (#07–2268, Millipore), anti-Atg14 (M184–3, MBL International, to detect human ATG14), anti-HA high affinity-HRP (#12013819001, Roche, to detect overexpressed TLR9-HA in cells and Tlr9-HA after immunoprecipitation from muscle lysates of the Tlr9-HA KI mice), anti-Flag M2-HRP (A8592, Sigma), and anti-GLUT4 (GT41-A, Alpha Diagnonstic International). To detect beclin 1 in Tlr9-HA immunoprecipitates from muscle lysates by western blot analysis, 10% MINI-PROTEAN TGX Precast gels (Bio-Rad) were used. For detection of all other proteins, 4–20% Precast gels were used.

Liu Y., Nguyen P.T., Wang X., Zhao Y., Meacham C.E., Zou Z., Bordieanu B., Johanns M., Vertommen D., Wijshake T., May H., Xiao G., Shoji-Kawata S., Rider M.H., Morrison S.J., Mishra P, & Levine B. (2020). Tlr9 and Beclin 1 Cross-Talk Regulates Muscle AMPK Activation in Exercise. Nature, 578(7796), 605-609.

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Comprehensive Antibody Collection for Cell Signaling

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Antibodies used in this study included anti-STING (13647; Cell Signaling Technology), anti-Flag M2 (F3165; Sigma-Aldrich), anti-HA (H3663; Sigma-Aldrich), anti-EGFP (GL-8; Clontech), anti-LC3 (7543; Sigma-Aldrich), anti-p62 (PM045; MBL), anti-STX17 (HPA001204; Sigma-Aldrich), anti-SNAP29 antibody (111303, SYSY, or sc-135564; Santa Cruz Biotechnology), anti-LAMP1(L1418; Sigma-Aldrich), anti-LAMP2 (sc-18822; Santa Cruz or L0668; Sigma-Aldrich), anti-Myc (9E10, DSHB), anti-WIPI2 (ab105459; Abcam), anti-FIP200 (17250; ProteinTech), anti-ATG16L (PM040; MBL), anti-α-Tubulin (E7; DSHB), anti-VAMP8 (ab76021; Abcam), anti-GABARAP (ab109364; Abcam), anti-AMPK (2532; Cell Signaling Technology), anti-p-AMPK T172 (2535; Cell Signaling Technology), anti-TBC1D1(66433; Cell Signaling Technology), anti-p-TBC1D1 Ser237(07-2268; Sigma-Aldrich), anti-Raptor (2280; Cell Signaling Technology), anti-p-Raptor Ser792 (2083; Cell Signaling Technology), anti-TSC2 (4308; Cell Signaling Technology), anti-p-TSC2 Ser1387 (5584; Cell Signaling Technology), anti-ACC (3662; Cell Signaling Technology), anti-p-ACC Ser79 (3661; Cell Signaling Technology), anti-Glut4 (GT-41-A; Alpha diagnostic international), anti-Laminin2 (L0663; Sigma-Aldrich), anti-CD36 (80080; Abcam), anti-Dystrophin (15277; Abcam), and LysoTracker (ENZ-51005; Enzo).

Rong Y., Zhang S., Nandi N., Wu Z., Li L., Liu Y., Wei Y., Zhao Y., Yuan W., Zhou C., Xiao G., Levine B., Yan N., Mou S., Deng L., Tang Z., Liu X., Kramer H, & Zhong Q. (2022). STING controls energy stress-induced autophagy and energy metabolism via STX17. The Journal of Cell Biology, 221(7), e202202060.

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Antibodies for Western Blot Analysis

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Western Blot Analysis of Neurodegeneration Markers

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Whole-cell extracts were resuspended in PBS in the presence of protease inhibitor and sonicated. 15 μg of protein sample were separated on a 10% Bis-Tris gel (Invitrogen) and transferred to a nitrocellulose membrane (Invitrogen). The membrane was blocked in 4% milk-PBST (PBS with 0.1% Tween-20) and proteins were stained using the following antibodies: anti-TDP-43 (rabbit, Proteintech, 1:1000), anti-GAPDH (rabbit, Proteintech, 1:1000), anti-HSP70 (rat, EnzoLife Science, 1:1000), anti-tubulin (mouse, available in house, 1:10,000) and HRP-conjugated secondary antibodies anti-rabbit (goat, Dako, 1:2000), anti-mouse (goat, Dako, 1:2000), anti-rat (rabbit, Dako, 1:2000).
To detect TBC1D1, 30 μg of protein sample were separated using a pre-cast gradient gel (4–12%, Invitrogen) and wet blotted onto a PVDF membrane (Merck Millipore). The membrane was then blocked in 4% milk-TBST, stained with anti-TBC1D1 (rabbit, Cell Signalling, 1:1 000, in 2% milk-TBST) and anti-rabbit HRP-conjugated secondary antibody (goat, Dako, 1:2 000).
Image acquisition and result quantification were conducted using Alliance Q9 Advanced Chemiluminescence Imager (UviTech).

Šušnjar U., Škrabar N., Brown A.L., Abbassi Y., Phatnani H., Cortese A., Cereda C., Bugiardini E., Cardani R., Meola G., Ripolone M., Moggio M., Romano M., Secrier M., Fratta P, & Buratti E. (2022). Cell environment shapes TDP-43 function with implications in neuronal and muscle disease. Communications Biology, 5, 314.

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