The following primary antibodies were used at the indicated dilutions: anti-LC3A/B (1:1000 for western blotting and 1:100 for immunofluorescence; Cell Signaling Technology, 4108), anti-LC3A (1:100 for immunohistofluorescence; Abgent, 1805a), anti-ATG12 (1:100; Cell Signaling Technology, 2010), anti-ATG13 (1:1000; Cell Signaling Technology, 6940), anti-ATG16L1 (1:1000; Cell Signaling Technology, 8089), anti-human LAMP1 and anti-mouse LAMP1 (1:100; Becton Dickinson, 555798 and 553792), anti-WIPI2 (1:100; Bio-Rad, MCA5780GA), anti-CDSN/corneodesmosin (R&D Systems, AF5725), anti-GAPDH (1:2000; Santa Cruz Biotechnology, sc-25778), and anti-ATP6V0D1 (1:50; ABCAM, ab56441). The following inhibitors and reagents were used: Amiodarone hydrochloride (Sigma, A8423), bafilomycin A1 (Tocris Biosciences, 1334), betahistine dihydrochloride (Sigma, B4638), chloroquine (Sigma, C6628), CCCP (Sigma, C2759), HBSS (Gibco, 14025–092), hydroxychloroquine (Sigma, H0915), lidocaine (Sigma, L7757), lidocaine hydrochloride monohydrate (Sigma, L5647), monensin (Sigma, M5273), NH4Cl (Sigma, 213330), nigericin (Sigma, N7143), PP242 (Tocris, 4257), procainamide hydrochloride (Sigma, P9391). Mito-TMRE (abcam, 113852), PIK3C3/VPS34 inhibitor (IN-1) was kindly provided by Dr I. Ganley, University of Dundee.
Procainamide hydrochloride
Manufactured by Merck Group
Sourced in United States
Procainamide hydrochloride is a chemical compound used in the production of various pharmaceutical and laboratory products. It is a white crystalline powder that is soluble in water and alcohol. Procainamide hydrochloride is commonly used as a raw material in the synthesis of other chemical compounds, but its specific core function is not available within the scope of this request.
Automatically generated - may contain errors
Lab products found in correlation
Dimethyl sulfoxide (dmso), by Merck Group (5 mentions)
Picoline borane, by Merck Group (3 mentions)
Ammonium hydroxide, by Merck Group (3 mentions)
Formic acid, by Merck Group (3 mentions)
Acetonitrile, by Merck Group (3 mentions)
Acetic acid, by Merck Group (3 mentions)
Pngase f, by New England Biolabs (3 mentions)
Nigericin, by Merck Group (1 mentions)
Amiodarone hydrochloride, by Merck Group (1 mentions)
Lidocaine hydrochloride monohydrate, by Merck Group (1 mentions)
Betahistine dihydrochloride, by Merck Group (1 mentions)
Hydroxychloroquine, by Merck Group (1 mentions)
Chloroquine, by Merck Group (1 mentions)
Monensin, by Merck Group (1 mentions)
Lidocaine, by Merck Group (1 mentions)
Anti gapdh, by Santa Cruz Biotechnology (1 mentions)
Anti lc3a b, by Cell Signaling Technology (1 mentions)
Anti atg12, by Cell Signaling Technology (1 mentions)
Anti atg16l1, by Cell Signaling Technology (1 mentions)
Anti atg13, by Cell Signaling Technology (1 mentions)
9 protocols using procainamide hydrochloride
1
Autophagy Protein Detection and Inhibition
2
Enzymatic Glycan Characterization
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Formic acid, ammonium-hydroxide, acetic acid, acetonitrile, picoline borane, procainamide-hydrochloride, and dimethyl sulfoxide were purchased from Sigma-Aldrich (St. Louis, MO, USA). PNGase F was provided by Asparia Glycomics (San Sebastian, Spain). An exoglycosidase sequencing kit was obtained from New England Biolabs (Ipswich, MA, USA).
3
Procainamide Labeling of Glycans
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ProA labeled glycans were prepared in the same way as 2-AB labeled glycans. The only difference was that the labeling mixture contained 38.3 mg/ml ProA (procainamide hydrochloride, Sigma-Aldrich) instead of 19.2 mg/ml 2-AB (equimolar concentrations).
4
N-Glycan Labeling and Purification
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N-glycans were released, fluorescently labeled, and cleaned up as described previously [25 (link)], with modification by using procainamide hydrochloride (ProA) as a fluorescent labeling agent for N-glycans. Briefly, isolated IgG was denatured with sodium dodecyl sulfate (SDS) (Invitrogen, USA) and by incubation at 65°C. After enzymatic release with PNGase F (Promega, USA), N-glycans were fluorescently ProA labeled in a 2-step procedure. First, 25 µL of procainamide mixture containing 4.32 mg of procainamide hydrochloride (Sigma-Aldrich) in glacial acetic acid (Honeywell, Charlotte, NC, USA)/dimethyl sulfoxide (Sigma-Aldrich) (30:70) was added per sample, followed by incubation at 65°C for 1 h. Then, 25 µL of reducing agent solution consisting of 4.48 mg of 2-picoline borane (J&K Scientific, Beijing, China) in glacial acetic acid/dimethyl sulfoxide (30:70) was added per sample and incubated at 65°C for an additional 1.5 h. Samples were then cooled for 30 min at RT. Excess reagents were removed from the samples using hydrophilic interaction liquid chromatography–solid phase extraction (HILIC-SPE) as described in [25 (link)]. For the HILIC-SPE step, an AcroPrepadv 1 mL 0.2 μm wwPTFE plate (Pall) was used as the stationary phase. The labeled glycans were eluted with ultrapure water and stored at − 20°C until use.
5
Biochemical Reagents Acquisition Protocol
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Formic acid, ammonium-hydroxide, acetic acid, acetonitrile, picoline borane, procainamide-hydrochloride, dimethyl sulfoxide, magnesium(II) nitrate hexahydrate and ethanolamine were purchased from Sigma-Aldrich (St. Louis, MO, USA). PNGase F was obtained from New England Biolabs (Ipswich, MA, USA). Nickel(II) nitrate hexahydrate and sodium acetate were obtained from Thermo Fisher Scientific (Kandel, Germany). Mangan(II) nitrate tetrahydrate was provided by Carl Roth (Karlsruhe, Germany). Cobalt(II) nitrate hexahydrate and iron(III) nitrate nonahydrate were purchased from VWR International (Leuven, Belgium).
6
Glycan Analysis Using PNGase F
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Formic acid, ammonium-hydroxide, acetic acid, acetonitrile, picoline borane, procainamide-hydrochloride and dimethyl sulfoxide were purchased from Sigma-Aldrich (St. Louis, MO, USA). Phosphate-Buffered Saline was obtained from VWR (Radnor, PA, USA). Ammonia solution was obtained from Scharlab S.L. (Barcelona, Spain). PNGase F was obtained from New England Biolabs (Ipswich, MA, USA).
7
Rapid PNGase F-Mediated Glycan Release
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All chemicals and reagents were at least analytical reagent grade and used without further purification. Rapid™ PNGase F and Rapid™ PNGase F Buffer were supplied by New England Biolabs (Ipswich, USA). Immunoglobulin from human serum (hIgG, ≥ 95%), human alpha-1-acid glycoprotein (hAGP, ≥ 95%), dextran Mw5000, dextran Mw1000, Discovery Glycan solid-phase extraction (SPE) tubes, and procainamide hydrochloride (≥ 98%) were purchased from Sigma-Aldrich (St. Louis, USA). Ammonium formate (> 99%) was obtained from VWR International (Radnor, USA). All solvents (acetonitrile, water) were LC–MS grade and purchased from Sigma-Aldrich (St. Louis, USA).
8
Insecticide Enzymes Purification Protocol
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Propoxur (99%) was purchased from Bayer Company Limited. Methomyl (80%) was from Shandong Jining Chemical Plant. Azamethiphos (95%) was from Shanghai Yongyuan Chemical Group Company Limited. Beta-cypermethrin (95.2%) and DDVP (98.7%) were obtained from Tianjin Longdeng Chemical Company Limited. Bifenthrin (95%) and Deltamethrin (99%) were from Jiangsu Yangnong Chemical Company Group Company Limited. Leupeptin, phenylmethanesulfonyl fluoride (PMSF), eserine hemisulfate, procainamide hydrochloride, bovine serum albumin (BSA), and acetylthiocholine iodide (ATChI) were acquired from Sigma Chemical Company. Tetraethylammonium iodide (Net4I) was from Sigma-Aldrich Company Limited. EDTA-Na2 was from BIO-RAD Laboratories. 2-Ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ) and 5,5’-dithiobis-(2-nitrobenzoic acid) (DTNB) were from Fluka Chemical Company. ECH-sepharose 4B was from Beijing Solarbio Science & Technology Company Limited. Sephadex G-25 columns were purchased from Amersham Pharmacia Biotech Incorporated. Other chemicals were purchased from commercial suppliers, all were analytical quality.
9
Glycoprotein Analysis Workflow
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Bovine
fetuin, human serum, trypsin (TPCK
treated), procainamide hydrochloride, and dimethyl sulfoxide (DMSO)
were purchased from Sigma. Ammonium bicarbonate, ammonium formate,
formic acid (for LC-MS), and sodium cyanoborohydride were purchased
from Fluka. PNGase F (Glycerol free) was purchased from New England
Biolabs (NEB). Sialidase S (recombinant from Streptococcus
pneumonia expressed in Escherichia
coli) was purchased from Prozyme. Acetonitrile (ACN,
HPLC grade) was purchased from Fisher. Octadecyl (C18) disposable
extraction columns were purchased from J.T. Baker. PD MiniTrap G-10
was purchased from GE Healthcare. Other reagents were analytical grade.
fetuin, human serum, trypsin (TPCK
treated), procainamide hydrochloride, and dimethyl sulfoxide (DMSO)
were purchased from Sigma. Ammonium bicarbonate, ammonium formate,
formic acid (for LC-MS), and sodium cyanoborohydride were purchased
from Fluka. PNGase F (Glycerol free) was purchased from New England
Biolabs (NEB). Sialidase S (recombinant from Streptococcus
pneumonia expressed in Escherichia
coli) was purchased from Prozyme. Acetonitrile (ACN,
HPLC grade) was purchased from Fisher. Octadecyl (C18) disposable
extraction columns were purchased from J.T. Baker. PD MiniTrap G-10
was purchased from GE Healthcare. Other reagents were analytical grade.
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