The following commercial antibodies were used: Huntingtin (MAB2166, 1:3000) and puromycin (MABE343, 1:10,000) antibodies were obtained from, Millipore-Sigma. Anti-polyglutamine (poly-Q) antibody (P1874, 1:5000) was from Sigma. Actin (sc47778, 1:20,000) and GST-horseradish peroxidase (HRP, sc138 HRP, 1:10,000) antibodies were from Santa Cruz Biotechnology. RPL7 (IHC-00455, 1:10,000), RPL35A (A305-106A, 1:10,000) and Caprin1 (A303-881A, 1:1000) from Bethyl Laboratories. mTOR (2972, 1:3000), FMRP (4317, 1:1500), and S6 (2217, 1:10,000), and normal mouse IgG (5415, 1:2500) were from Cell Signaling Technology. Mfsd (10 11518-1-527 AP, 1:1000), Acan (13880-1-AP, 1:1000), Ppbp (13313-1-AP, 1:1000), Mgp (10734-1-AP, 1:1000), and Phf11d (10898-1-AP, 1:1000) were from Proteintech. HRP-conjugated secondary antibodies were from Jackson ImmunoResearch Inc. HRP-conjugated secondary antibodies (115-035-146 (goat anti-mouse), 1:10,000; or 111-035-144 (goat anti-rabbit), 1:10,000) were from Jackson ImmunoResearch Inc. For Immunostaining Huntingtin (1:100, MAB2166), Rpl7 (1:50; IHC-00455) were used. Secondary antibodies for STED microscopy were anti-mouse STAR 635p (1:400) and anti-rabbit Alexa 594 (1:400) that were self-coupled in-house and the company that produced them is Abberior, Göttingen, Germany.
Mtor 2972
Manufactured by Cell Signaling Technology
Sourced in United States
The MTOR (#2972) is a laboratory instrument used to detect and measure the phosphorylation of the mammalian target of rapamycin (mTOR) protein. mTOR is a serine/threonine protein kinase that plays a central role in regulating cell growth, proliferation, and metabolism. The MTOR (#2972) can be used to analyze mTOR signaling and activity in various cell and tissue samples.
Automatically generated - may contain errors
Lab products found in correlation
S6 2217, by Cell Signaling Technology (1 mentions)
Acan 13880 1 ap, by Proteintech (1 mentions)
Mgp 10734 1 ap, by Proteintech (1 mentions)
Actin sc 47778, by Santa Cruz Biotechnology (1 mentions)
Mab2166, by Merck Group (1 mentions)
Hrp conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions)
Enhanced chemiluminescence western blotting substrate, by Santa Cruz Biotechnology (1 mentions)
Gapdh sc 32233, by Santa Cruz Biotechnology (1 mentions)
Bca protein assay reagent, by Thermo Fisher Scientific (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Ab32047, by Abcam (1 mentions)
P mtor 5536, by Cell Signaling Technology (1 mentions)
Tfeb 13372 1 ap, by Proteintech (1 mentions)
Goat anti rabbit and anti mouse igg hrp antibodies, by Bioworld Technology (1 mentions)
Chloroquine cq, by Merck Group (1 mentions)
Tert butyl hydroperoxide solution tbhp, by Merck Group (1 mentions)
Collagenase type 2, by Merck Group (1 mentions)
Hematoxylin eosin staining kit, by Solarbio (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
β actin bs 0061r, by Bioss Antibodies (1 mentions)
5 protocols using mtor 2972
1
Antibody Characterization for Huntington's Disease Research
2
Protein Extraction and Western Blot Analysis
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Total protein was isolated from tissues using RIPA buffer contains 50 mM Tris-HCl (pH 8.0), 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate and 0.1% SDS. Protein concentrations were measured using BCA Protein Assay Reagent (Thermo scientific). Proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), transferred to a polyvinylidene fluoride (PVDF) membrane (Millipore Corporation), blocked in 5% fat-free milk for 1 h at RT, then incubated with first antibodies (diluted in 5% milk) overnight at 4 °C. Lkb1 (sc-32245, 1:1000, RRID: AB_6278 ) and GAPDH (sc-32233, 1:1000, RRID: AB_627679 ) antibodies are from Santa Cruz biotechnology. mTOR (#2972, 1:1000, RRID: AB_330978 ) antibodies are from Cell signaling technology. The horseradish peroxidase (HRP)-conjugated secondary antibody (anti-rabbit IgG, 111-035-003, RRID: AB_10015289 or anti-mouse IgG, 115-035-003, RRID: AB_10015289 , Jackson ImmunoResearch) were diluted 1: 10,000. Immunodetection was performed using enhanced chemiluminescence western blotting substrate (Santa Cruz biotechnology) and detected by FluorChem R System (ProteinSimple). Results shown in the figures are representative results from at least three independent experiments.
3
Chondroprotective Effects of Apigenin
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Apigenin (C15H10O5; purity ≥98%), hematoxylin–eosin (HE) staining kit, and Safranin O-fast green (SO) staining kit were purchased from Beijing Solarbio Science & Technology (Beijing, China); tert-butyl hydroperoxide solution (TBHP), chloroquine (CQ), and type II collagenase were obtained from Sigma-Aldrich (St. Louis, MO, United States); dorsomorphin (compound C) and bafilomycin A1 were purchased from MCE (Monmouth Junction, NJ, United States); the primary antibodies against Bcl2 (12789-1-AP), Bax (50599-2-Ig), MMP13 (18165-1-AP), β-actin (20536-1-AP), and TFEB (13372-1-AP) were obtained from Proteintech (Wuhan, China); p21 (ab109199), p62 (ab109012), collagen II (ab188570), ADAMTS5 (ab41037), AMPK (ab32047), p-AMPK (ab13448), LAMP2 (ab125068), and lamin B (ab133741) were acquired from Abcam; LC3 (#83506), cleaved caspase 3 (#9661), p-mTOR (#5536), and mTOR (#2972) and apigenin were partially purchased from Cell Signaling Technology (Danvers, MA, United States); p16INK4a (A0262), aggrecan (A11691), and LAMP1 (A2582) were purchased from ABclonal Technology (Wuhan, China). Goat anti-rabbit and anti-mouse IgG-HRP antibodies were purchased from Bioworld (Nanjing, China). Alexa Fluor® 594- and 488-conjugated secondary antibodies were obtained from Jackson ImmunoResearch (PA, United States); 4′,6-diamidino-2-phenylindole (DAPI) was purchased from Beyotime (Shanghai, China).
4
Icariin Modulates Bone Metabolism
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Icariin (20200719) was purchased from Xi'an Yuhui Biotechnology Co. Ltd (Xi'an, China). Alkaline phosphatase (ALP) (JL26470), procollagen type I N-terminal propeptide (PINP) (JL49448), tartrate-resistant acid phosphatase isoform 5b (TRACP-5b) (JL12318), and C-telopeptide of type I collagen (CTX-I) (JL20748) enzyme-linked immunosorbent assay (ELISA) kits were purchased from Jianglai (Shanghai, China). Autophagy-related protein 7 (Atg7) (8558), Akt (9272), p-Akt (4058), Beclin1 (3495), mammalian target of rapamycin (mTOR) (2972), p-unc-51-like autophagy activating kinase 1 (ULK1) (Ser555) (5869), p-mTOR (2971), ULK1 (8054), p-ULK1 (Ser757) (14202), and AMPactivated protein kinase (AMPK) (5832) were purchased from Cell Signaling Technology (Danvers, USA). Microtubule-associated protein 1 light chain 3 (LC3) (GTX127375) was purchased from Genetex (Louis Park, USA), sequestosome 1 (p62) (ab109012) from Abcam (Cambridge, UK), β-actin (bs-0061R) from Bioss (Woburn, USA) and p-AMPK (13S4010) from Bioword (Louis Park, USA). Bicinchoninic acid (BCA) kits were purchased from Thermo Fisher Scientific (Waltham, USA) and dithiothreitol (DTT; 1 mol/L) was obtained from Sigma-Aldrich (St. Louis, USA).
5
Sirolimus-Based Polydimethylsiloxane Formulation
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Sirolimus (rapamycin, R-5000) was purchased from LC Laboratories (Woburn, MA, USA). Polydimethylsiloxane (PDMS; Sylgard® 184 silicone elastomer kit) was purchased from Dow Corning (Midland, MI, USA). Rhodamine B (RhoB, R6626) and Dulbecco’s phosphate-buffered saline (D8662) were purchased from Sigma Aldrich (St. Louis, MO, USA). Dimethyl sulfoxide (D0457) was purchased from Samchun Chemical (Seoul, Korea). Zoletil® 50 was purchased from VIRBAC (Virbac, Carros, France), and heparin sodium was obtained from Hanlim (Seoul, Korea). Aspirin was purchased from BayerKorea (Seoul, Korea), and clopidogrel bisulfate was purchased from Plavix (Sanofi Winthrop Industries, Paris, France). Solvents for high-performance liquid chromatography were purchased from Merck Inc. (Darmstadt, Germany). All chemicals and reagents were of analytical grade. Radioimmunoprecipitation assay (RIPA) lysis buffer (R0146CD) was obtained from Bylabs (Hanam, Korea). mTOR (#2972) and phospho-mTOR (Ser2248, #2971) were purchased from Cell Signaling Technology (Beverly, MA, USA). Proliferation cell nuclear antigen (PCNA) (ab19166), α-SMA (ab5694), LC3B (ab48394), and β-actin (mAbcam 8224) were purchased from Abcam (Cambridge, MA, USA).
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