TSC1f/f-ERCre+ mice were previously reported [27 (link), 56 (link)]. LysM-Cre+ mice were purchased from Jackson Laboratory. M. bovis (Bacillus Calmette-Guérin or BCG) was obtained from the laboratory of Dr. William Jacobs (Albert Einstein College of Medicine). Virulent M. tuberculosis H37Rv (ATCC, 25618D-2) was purchased from ATCC. Rapamycin was obtained from Enzo Life Science (Enzo Life Science, A275). Bafilomycin A1 (B1793), 3-methyladenine (3-MA, M9281), and wortmannin (W1628) were purchased from Sigma-Aldrich. Compound C (Millipore, 171,260) was obtained from Millipore (Billerica, MA).
Wortmannin w1628
Manufactured by Merck Group
Sourced in United States
Wortmannin (W1628) is a chemical compound that inhibits the phosphoinositide 3-kinase (PI3K) enzyme. It is commonly used as a research tool in cell biology and biochemistry studies.
Automatically generated - may contain errors
Lab products found in correlation
3 ma m9281, by Merck Group (2 mentions)
Rapamycin, by Enzo Life Sciences (1 mentions)
Bafilomycin a1 b1793, by Merck Group (1 mentions)
Compound c, by Merck Group (1 mentions)
Lysm cre mice, by Jackson ImmunoResearch (1 mentions)
Bafilomycin a1, by Enzo Life Sciences (1 mentions)
Ku 60019, by Selleck Chemicals (1 mentions)
Lm001 11, by Welgene (1 mentions)
Lm001 79, by Welgene (1 mentions)
Torin 1, by Biorbyt (1 mentions)
Hrp conjugated goat anti rabbit igg h l, by Jackson ImmunoResearch (1 mentions)
Monoclonal anti flag hrp, by Merck Group (1 mentions)
P70 s6 kinase, by Cell Signaling Technology (1 mentions)
Myc tag 71d10 rabbit mab, by Cell Signaling Technology (1 mentions)
Hrp conjugated goat anti mouse igg h l, by Jackson ImmunoResearch (1 mentions)
Phospho 4e bp1 thr37 46, by Cell Signaling Technology (1 mentions)
Polyclonal anti lc3, by Merck Group (1 mentions)
Alexa fluor 555 goat anti mouse, by Thermo Fisher Scientific (1 mentions)
Polyclonal anti actin, by Merck Group (1 mentions)
Monoclonal anti p62, by Santa Cruz Biotechnology (1 mentions)
10 protocols using wortmannin w1628
1
Genetic Murine Models in Tuberculosis Research
2
Senescent Fibroblast Induction Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Normal human fibroblasts isolated from healthy newborn foreskins were provided by Dr. Jin-Ho Chung (IRB No. H-1101-116-353 from the School of Medicine, Seoul National University, Korea) and maintained in Dulbecco’s modified Eagle’s medium (DMEM) (LM001-11; Welgene, Korea) supplemented with 10% fetal bovine serum (S-FBS-US-015; Serana, Australia) at 5% CO2 and 37°C. The DMEM contained no glucose (LM001-79; Welgene) to simulate glucose deprivation. Cells were passaged at a 1:4 dilution until they stopped dividing at passage 32 to obtain a fibroblast population at replicative senescence. Fibroblasts mid-passage (between 20 and 24) were pulsed with 0.25 μM doxorubicin for 4 h and chased in fresh medium replaced every two days for chemically induced senescence. On Day 5, cells began expressing senescence phenotypes similar to previous studies (Song et al., 2005 (link)). Chemicals purchased from the following sources were used at 200 nM, 250 nM, 50 μM, 0.25 μM, 0.2 μM, and 0.5 μM doses, respectively: bafilomycin A1 from Enzo Life Science (USA) (BML-CM110-0100); Torin-1 from Biorbyt (UK) (orb146133); chloroquine (CQ; C6628), doxorubicin (D1515), and wortmannin (W1628) from Sigma-Aldrich (USA); and KU60019 from Selleckchem (USA) (S1570).
3
Antibody Profiling for Autophagy Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We used the following primary antibodies: monoclonal antibodies (mAbs) against LC3 (M186-3) and ATG7 (PM039) from MBL, phospho-p70S6 kinase (#9206), p70S6 kinase (Thr389, #9202), phospho-4E-BP1 (Thr37/46, #9459), Myc-Tag (71D10) rabbit mAb (#2278 s) from Cell Signaling Technology. Rac1-102 mAb (#610650) from BD Transduction Laboratories was used to specifically detect the non-glucosylated Rac1 (Non-glu. Rac1), and Rac1 mAb 23A8 (#05-389) from Millipore was used to detect the total Rac1. Monoclonal anti-Flag-HRP (#A8592) was ordered from Sigma. The secondary antibodies HRP-conjugated goat anti-mouse IgG (H+L) (#115-035-003) and HRP-conjugated goat anti-rabbit IgG (H+L) (#111-035-003) were purchased from Jackson Immuno Research. The following chemicals were used: chloroquine (CQ), 3-Methyladenine (3-MA, M9281) and wortmannin (W1628) from Sigma, and 3-90 -2,5-diphenyltetrazolium bromide (MTT) from Amresco.
4
Autophagy Regulation by QSOX1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell culture reagents were purchased from Invitrogen. Earle’s balanced salt solution (EBSS, E3024), bafilomycin A1 (B1793), 3-methyladenine (3-MA) (M9281) and wortmannin (W1628) were purchased from Sigma Aldrich. For the western blotting experiments, the following antibodies were used: polyclonal anti-rat QSOX1 [5] (link) diluted at 1∶7500, polyclonal anti-LC3 (Sigma, L8918) diluted at 1∶3000, monoclonal anti-p62 (Santa Cruz, sc-28359) diluted at 1∶1500, polyclonal anti-actin (Sigma, A5060) diluted at 1∶15000, polyclonal anti-rabbit (P.A.R.I.S, BI2407) diluted at 1∶10000 and polyclonal anti-mouse (P.A.R.I.S, BI24130) diluted at 1∶10000. For the immunofluorescence experiments, the following antibodies were used: monoclonal anti-p62 antibody (Santa Cruz, sc-28359) diluted at 1∶250, monoclonal anti-mouse lysosomal-associated membrane protein 1 (LAMP1) (Abcam, Ab25630) diluted at 1∶100, Alexa Fluor 555 goat anti-mouse (Life technologies, A-21422) diluted at 1∶800.
5
RAW 264.7 Cell Culture Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RAW 264.7 cells were from ATCC (ATCC TIB‐71) and were tested several times for mycoplasma contamination. They were cultured in DMEM medium supplemented with 10% FBS, 4.5 g/l of glucose, 2 mM of Glutamine, and 100 U/ml of Penicillin/Streptomycin. LPS was purchased from Sigma (L4391) and mouse IFNγ, IL‐4, and IL‐13 were from R&D Systems (485‐MI, 404‐ML, and 413‐ML). Wortmannin (W1628) and AS1517499 (SML1906) were purchased from Sigma. Flag‐Ric8b WT and Flag‐Ric8b MUT were generated by VectorBuilder using the pLV‐Puro‐CMV/T7‐EGFP as a lentiviral vector backbone.
6
Immunofluorescence Labeling of Endosomal Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary antibodies used were as follows: rabbit polyclonal anti-Tsg101 (#ab30871, Abcam, Cambridge, MA, USA), rabbit polyclonal anti-CD63 (#sc-15363, Santa Cruz Biotechnology, CA, USA), mouse monoclonal anti-ubiquitin (#MK-11-3, MBL, Woburn, MA, USA), mouse monoclonal anti-LBPA (#Z-PLBPA, Echelon, Salt Lake City, UT, USA), and rat monoclonal anti-E-cadherin (#U3254, Sigma-Aldrich, St. Louis, MO, USA). Secondary antibodies used for immunofluorescence staining are as follows: chick anti-rabbit Alexa Fluor 488, donkey anti-mouse Alexa Fluor 568, and goat anti-rat Alexa Fluor 647 antibodies (Invitrogen, Carlsbad, CA, USA). To counterstain nuclei, TO-PRO-3-iodide (Invitrogen) or SYTO 11 green fluorescent nucleic acid stain (Invitrogen) was used. For confocal live imaging of MVBs, 1,1′-Dioctadecyl-3,3,3′,3′ -tetramethylindocarbocyanine iodide (Di-I) (AS-84711; ANASPEC, Fremont, CA, USA) (absorption/emission at 549/565 nm). PD173074, a FGFR antagonist, was purchased from Selleckchem (S-1264, Houston, TX, USA), and wortmannin (W-1628) and and 3-MA (M-9281) were purchased from Sigma-Aldrich.
7
Oxidative Stress and Autophagy Modulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
P4 (P0130) and 17β-estradiol (E2, E8875) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Sesame oil (241002500) was purchased from Acros Organics (Leicestershire, UK). Three-methyladenine (3-MA) (M9281) and wortmannin (W1628) were purchased from Sigma-Aldrich. LysoTracker Red DND-99 (L7528), MitoTracker Red CMXRos (M7512), and non-fluorescent compound 5-(and-6)-chloromethyl-2', 7'-dichlorodihydrofluorescein diacetate, acetyl ester (CM-H2DCFDA) (C6827) were purchased from Invitrogen (Carlsbad, CA, USA). H2O2 was purchased from Sigma-Aldrich.
8
Estrogen Receptor Signaling Modulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibodies against β-actin (no. 4967), Phospho-Akt (Ser473) (no.9271) were purchased from Cell Signaling Technology, Inc. (Danvers, MA). Anti-C10orf58 (PAMM) antibody (HPA009025), 17β-estradiol (E2) (E8515), corn oil (C8267), H2O2 (H1009). Wortmannin (W1628) and Rapamycin (R8781) were purchased from Sigma-Aldrich Inc. (St. Louis, MO). The high affinity estrogen receptor antagonist ICI 182,780 was purchased from TOCRIS Bioscience (Minneapolis, MN-Cat no. 1047).
9
Regulated Cell Adhesion Disruption
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MDCK, A431D, and DLD1 cells (American Type Culture Collection) and DLD1 R2/7 αCat-deficient colon carcinoma cells (a gift from F. van Roy, Ghent University, Ghent, Belgium) were maintained in DMEM (Corning), containing 10% FBS (Atlanta Biologicals or JRS Scientific), 100 U/ml penicillin, and 100 µg/ml streptomycin (Corning). iDimerize cell lines were sorted for positive selection of mCherry fluorescence by flow cytometry using FACSAria 5 (BD Biosciences). mCherry FLαCat or FLαCatKKR<3A cell lines were selected in 5 µg/ml puromycin (Sigma-Aldrich) and were subsequently sorted for mCherry expression using flow cytometry. In drug treatment assays, cells were treated with one of the following: DMSO control (Sigma-Aldrich) or 1–20 µM wortmannin (W1628; Sigma-Aldrich) for 3 h. In forced-dimerization assays, cells were treated with 25, 250, or 500 nM B/B homodimer (635059; Clontech) for 20 min to 15 h, which was sometimes followed by treatment with 1 µM washout ligand (635088; Clontech).
10
Antibody-based Immunoblotting Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rabbit anti-LC3A/B (4108), anti-p-AKT (Ser473) (4060), anti-AKT (4691), and anti-p-MTOR (Ser2448; 5536), anti-MTOR (2983), antipAMPK (4185), anti-AMPK (2532), anti-p-ERK (4370), anti-ERK (4695), antip-P38 (4511), and anti-P38 (9212) antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA). Rabbit anti-P62 (3340-1) antibodies were purchased from Epitomics (Burlingame, CA, USA). Rabbit anti-CASP2 (ab179520), anti-ULK1 (ab128859), and anti-ATG5 (ab108327) antibodies were purchased from Abcam (Cambridge, MA, USA). Mouse anti-ATG7 (sc-376212) and anti-BECN1 (sc-48341) antibodies was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Mouse anti-Flag (clone M2) (F1804) mAb, wortmannin (W1628), and 3-MA (M9281) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Rabbit anti-MYC (R12081) and anti-GAPDH/glyceraldehyde-3-phosphate dehydrogenase (EM1101) antibodies were purchased from Huaan Biological Technology (Hangzhou, China). Mouse monoclonal antibodies (mAbs) recognizing theN/P proteinsof RABV were produced in our laboratory [15] .
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!