Lysates and eluates from Immunoprecipitations were run on 10% acrylamide gels for SDS–PAGE, freshly prepared and used the same day: 10% Protogel (30% w/v acrylamide, 0.8% bis‐acrylamide (37.5:1 solution, National diagnostics, Atlanta, USA), 380 mM Tris–HCl pH 8.8, 0.1% w/v SDS (Applichem, Darmstadt, Germany), 0.06% v/v TEMED (Applichem), 0.06% w/v APS (Applichem) for the running gel and 5% Protogel, 165 mM Tris–HCl pH 6.8, 0.1% w/v SDS, 0.08% v/v TEMED, 0.04% w/v APS for the stacking gel. Running buffer for SDS–PAGE was 190 mM glycine (Applichem), 25 mM Tris‐base (Applichem), 0.5% SDS. To facilitate Atg18 migration and avoid formation of aggregates, samples were reduced and denatured at 90°C using NuPAGE buffer (Thermo Fisher) containing LDS instead of SDS and supplemented with 100 mM DTT. Gels were blotted on 0.45 µm nitrocellulose membrane (Amersham) overnight at a constant current of 200 mA using a Trans‐Blot® Cell (Bio‐Rad, USA). Membranes were decorated using anti‐mCherry‐1C51 (Abcam), anti‐HA.11‐16B12 (BioLegend), anti‐G6PDH (Sigma‐Aldrich), anti‐Tubulin (clone B5‐1‐2, Sigma‐Aldrich), and anti‐WIPI1 (C‐terminal epitope, Sigma‐Aldrich).
Glycine
Manufactured by AppliChem
Sourced in Germany
Glycine is a pure amino acid commonly used in laboratory settings. It serves as a fundamental building block for various biochemical reactions and analyses. This product provides a reliable and consistent source of glycine for researchers and scientists.
Automatically generated - may contain errors
Lab products found in correlation
Tween 20, by Carl Roth (2 mentions)
Tris ultrapure, by AppliChem (2 mentions)
Am630, by Cayman Chemical (2 mentions)
Iron chloride hexahydrate, by Merck Group (2 mentions)
Rutin quercetin 3 o rutinoside hydrate, by Merck Group (2 mentions)
Gallic acid hydrate, by ITW Reagents (2 mentions)
Folin ciocalteu reagent, by Merck Group (2 mentions)
Orthovanadate, by Merck Group (2 mentions)
Phenylmethylsulfonyl fluoride pmsf, by Merck Group (2 mentions)
Luminol, by Merck Group (2 mentions)
Triton x 100, by Merck Group (2 mentions)
Capsazepine, by Merck Group (2 mentions)
Fetal calf serum (fcs), by PAN Biotech (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Lipofectamine rnaimax transfection reagent, by Thermo Fisher Scientific (2 mentions)
Nitrocellulose membrane, by Cytiva (1 mentions)
Trans blot cell, by Bio-Rad (1 mentions)
Bis acrylamide, by National Diagnostics (1 mentions)
Anti g6pdh, by Merck Group (1 mentions)
Protogel, by National Diagnostics (1 mentions)
11 protocols using glycine
1
Immunoprecipitation and SDS-PAGE Analysis
2
Immunostaining of HCV NS5A Protein
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Cells were fixed with 4% para-formaldehyde (PFA) 72 hpi, washed with 100 mM Glycine (Applichem), semi-permeabilized with 0.1% Triton X-100 (Vetec Labs) and stained for NS5A using sheep anti-NS5A61 (link) and Alexa Fluor anti-sheep 594 secondary antibody. Infectivity was expressed as focus-forming units per milliliter of supernatant (FFU/mL).
3
VEGF Signaling Pathway Assay
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AM630 (#Cay10006974) was obtained from Cayman Chemical (Ann Arbor, MI, USA). Recombinant human VEGF-165 (rVEGF, #HZ-1038) and recombinant human serum albumin (rHSA, #HZ-3001) were purchased from Proteintech (Planegg-Martinsried, Germany). Dimethyl sulfoxide (DMSO), glycerin, glycine, sodium chloride (NaCl), sodium hydroxide (NaOH), hydrochloric acid 37% (HCl), Tris hydrochloride (Tris-HCl), and Tris ultrapure were purchased from AppliChem (Darmstadt, Germany). β-Mercaptoethanol was purchased from Ferak (Berlin, Germany). Aqua ad iniectabilia was bought from Braun Melsungen (Melsungen, Germany). (-)-trans-Caryophyllene (BCP; #22075), hydrogen peroxide solution (H2O2), luminol, bromophenol blue, p-coumaric acid, and paraformaldehyde (PFA) were purchased from Sigma-Aldrich (Taufkirchen, Germany). Acrylamide (Rotiphorese® Gel 30), ammonium peroxydisulphate (APS), N,N,N′,N′-tetramethylethylenediamine (TEMED), and Tween® 20 were purchased from Carl Roth (Karlsruhe, Germany). Dulbecco’s phosphate-buffered saline (DPBS) and fetal bovine serum (FBS) were purchased from PAN Biotech (Aidenbach, Germany). Non-fat milk (NFM) powder was obtained from Bio-Rad Laboratories (Munich, Germany). GibcoTM Penicillin-Streptomycin (10,000 U/mL), GibcoTM Trypsin-EDTA, and GibcoTM Trypan Blue Solution were purchased from Thermo Fisher Scientific (Schwerte, Germany).
4
Biotinylation and Isolation of Cell Surface ADAM17
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ADAM17 expression in the DLD‐1 A17wt, I762A, and LEE cell lines was induced and 24 h later, cells were washed twice in cold PBS and incubated for 30 min with 0.5 mg/ml non‐cleavable EZ‐Link Sulfo‐NHS‐LC‐Biotin (Thermo Scientific) in PBS. The biotinylation was quenched by washing three times with 100 mM glycine (AppliChem) in PBS and additionally three PBS washes. Cells were lysed for 30 min in RIPA buffer supplemented with protease inhibitors. Cell lysates were cleared by centrifugation, protein concentration equalized by BCA assay, and supernatants incubated with streptavidin‐agarose beads (Sigma‐Aldrich) for 2 h at 4°C. Beads were washed three times in RIPA and bound proteins released by heating 5 min at 95°C in 2× SDS–PAGE sample buffer. Samples were analyzed by Western blot.
5
Quantification of Phenolic Compounds
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Gallic acid hydrate was from Panreac (Barcelona, Spain). Glycine (99.5%) was from Applichem (Darmstadt, Germany). Iron chloride hexahydrate was from Merck (Darmstadt, Germany). L-Lactic acid, ascorbic acid (99.5%), sodium acetate trihydrate, sodium carbonate anhydrous (99%) and aluminum chloride anhydrous (98%) were from Penta (Praha, Czechia). Chlorogenic acid (95%) was from Fluorochem (Hadfield, UK). NeoChlorogenic acid (≥98%) was from Merck (Darmstadt, Germany). Folin-Ciocalteu reagent, rutin (quercetin 3-O-rutinoside) hydrate (>94%), isorhamnetin 3-O-glucoside and 2,4,6-tris(2-pyridyl)-s-triazine (TPTZ) were from Sigma-Aldrich (St. Louis, MO, USA).
6
Crosslinked Microcarriers for Chondrogenesis
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Preparation of crosslinked microcarriers was performed as already described in our previous work16 (link). Briefly, microcarriers were incubated for 2 h at 37°C in 160 µM RB solution (AppliChem), followed by 250 mJ/sec UV-A irradiation for 5 min into a UV crosslinker (Stratalinker). TGF-β1 (Peprotech) was covalently bound to activated microcarriers for 2 h at room temperature in a final volume of 500 µL, and then quenched with 50 mM glycine (AppliChem). Crosslinked microcarriers were used as a support for chondrogenic differentiation. Cell attachment to scaffolds was promoted by incubating cell suspension with microcarriers in non-adherent propylene tubes (Falcon; Faust, Schaffhausen, Switzerland).
7
Antioxidant Compounds Characterization
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Glycine (99.5%) was from Applichem (Darmstadt, Germany). Iron chloride hexahydrate was from Merck (Darmstadt, Germany). Rutin (quercetin 3-O-rutinoside) hydrate, kaempferol 3-O-glucoside, 2,4,6-tris(2-pyridyl)-s-triazine (TPTZ), 2,2-diphenylpicrylhydrazyl (DPPH) and Folin-Ciocalteu reagent were from Sigma-Aldrich (St. Louis, MO, USA). L-Lactic acid, sodium carbonate anhydrous (99%), ascorbic acid (99.5%), and sodium acetate trihydrate and aluminium chloride anhydrous (98%) were from Penta (Praha, Czechia). Gallic acid hydrate was from Panreac (Barcelona, Spain). Pelargonin (pelargonidin 3,5-di-O-glucoside) chloride was from Extrasynthese (Genay, France).
8
Cannabinoid Receptor Signaling Pathway
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THC was obtained from Lipomed (Weil am Rhein, Germany). O-1602 and PD98059 were bought from R&D Systems (Wiesbaden-Nordenstadt, Germany). AM-251 and AM-630 were purchased from Biomol GmbH (Hamburg, Germany). Dulbecco's Modified Eagle's medium (DMEM, high glucose, GlutaMAXTM), trypsin-EDTA and penicillin-streptomycin were obtained from Life Technologies GmbH (Darmstadt, Germany). Fetal calf serum (FCS) was from PAN Biotech (Aidenbach, Germany). Dimethyl sulfoxide (DMSO), EDTA, glycerol, glycine, H2O2, HEPES, MgCl2, NaCl, sodium dodecyl sulphate and Tris were bought from AppliChem GmbH (Darmstadt, Germany). Aprotinin, capsazepine, HCl, leupeptin, luminol, orthovanadate, para-cumaric acid, phenylmethylsulfonyl fluoride (PMSF) and Triton® X-100 were obtained from Sigma-Aldrich (Taufkirchen, Germany). Milk powder was obtained from Bio-Rad Laboratories GmbH (Munich, Germany). Tween® 20 was purchased by Carl Roth GmbH (Karlsruhe, Germany).
9
Mifepristone-Induced VSV-G Protein Localization
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BHK-G43 cells were seeded into 24-well plates containing 12-mm glass coverslips and cultured for 24 h with GMEM containing 5% FBS. The nearly confluent cells received fresh cell culture medium containing mifepristone (10−9 M) and either NH125 (1–10 µM), bafilomycin A1 (0.08 µM), or DMSO (0.1%, v/v), and were maintained for 24 h at 37 °C with 5% CO2. The cells were washed once with PBS (4 °C) and fixed for 30 min at room temperature with 3% paraformaldehyde (AppliChem, Darmstadt, Germany). The cells were washed twice with PBS containing 0.1 M glycine (AppliChem) and once with PBS. Subsequently, the cells were incubated with a mouse monoclonal antibody (hybridoma clone I1, American Type Culture Collection, Manassas, VA, USA) directed to the VSV-G protein (1:50), washed three times with PBS, and then incubated for 1 h at room temperature with a goat anti-mouse IgG AlexaFluor-488 conjugate (1:500, Life Technologies Europe). The cells were rinsed twice with PBS and once with distilled water and the nuclei stained for 5 min with 0.1 µg/mL of diamidino-2-phenylindole (DAPI; Sigma) dissolved in ethanol. Finally, the cells were washed with distilled water and embedded in Mowiol 4–88 (Sigma) mounting medium.
10
Inducing NMDAR-Dependent Structural Plasticity
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To investigate structural synaptic plasticity, NMDA receptor dependent LTP was induced chemically (cLTP) by glycine. For this purpose, transfected cultures were incubated in 500 mL of Hank's balanced salt solution (HBSS 1X, GIBCO) for 10 min at room temperature followed by another 10 min incubation in HBSS 1X without magnesium containing 200 mM glycine (AppliChem) and 3 mM Strychnine (blocker for inhibitory ionotropic glycine receptor) (Sigma-Aldrich) (Fortin et al., 2010) . Afterward, the solution was removed and 500 mL of HBSS 1X per well were added and cells were incubated for another 50 min at room temperature before fixation. Control wells were treated only with HBSS 1X. Finally, cells were fixed using 4% paraformaldehyde (PFA) for 15 min at room temperature. The cultures were washed three times (each time 5 min) with PBS 1X. After washing, the coverslips were mounted using Fluoro-gel mounting medium (Electron Microscopy Sciences, Hatfield, PA).
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