Lipopolysaccharide (LPS) from E. coli 055: B5, LC–MS and HPLC-grade solvents and fine chemicals were procured from Sigma-Aldrich, USA. SYBR green mix cDNA synthesis kits were purchased from Takara Bioscience, India. ELISA kits (IL-6, CCL2, INF-γ and IL-1β) were obtained from R&D Systems. Primary antibodies were purchased from Cell Signaling Technology, USA, and the corresponding catalogue numbers are mentioned in supplementary table S2. Bicinchoninic acid reagent (BCA kit) was bought from Thermo Scientific, USA. Polyvinylidene difluoride (PVDF) membrane (0.45 μm) was procured from Millipore, USA. Secondary antibodies were procured from Jackson Laboratory, USA, and an ECL kit was purchased from Advansta, Menlo Park, CA, USA. Deionized water (18 MΩ) was used for the preparation of all solutions. Leaves of G. Sylvestre were collected from Chittoor district, Andhra Pradesh, India. The plant specimen was authenticated by a botany professor from Osmania University, Hyderabad, India. LC–MS CHROMASOLVs-grade and HPLC-grade methanol (MeOH) and acetonitrile (ACN) were procured from Sigma-Aldrich (St Louis, USA). Analytical reagent (AR)-grade ammonium acetate, ammonium formate, formic acid, hydrochloric acid (HCl).
Polyvinylidene difluoride pvdf membrane 0.45 μm
Manufactured by Merck Group
Sourced in United States
Polyvinylidene difluoride (PVDF) membrane (0.45 μm) is a laboratory filtration material composed of a semi-crystalline thermoplastic polymer. It has a pore size of 0.45 micrometers and is commonly used for filtration and separation applications in research and analytical settings.
Automatically generated - may contain errors
Lab products found in correlation
Ecl kit, by Advansta (1 mentions)
Lipopolysaccharide lps from e coli 055 b5, by Merck Group (1 mentions)
Acetonitrile acn, by Merck Group (1 mentions)
Inf γ, by R&D Systems (1 mentions)
Il 1β, by R&D Systems (1 mentions)
Urea, by Bio-Rad (1 mentions)
Iodoacetic acid, by Merck Group (1 mentions)
Thiourea, by Bio-Rad (1 mentions)
Mineral oil, by Bio-Rad (1 mentions)
Sodium dodecyl sulfate (sds), by Bio-Rad (1 mentions)
Bis acrylamide, by Merck Group (1 mentions)
Pierce 660 nm protein assay kit, by Thermo Fisher Scientific (1 mentions)
Proteomics grade trypsin from porcine pancreas, by Merck Group (1 mentions)
Chaps 3 3 cholamidopropyl dimethylammonio 1 propanesulfonate, by Bio-Rad (1 mentions)
Anti age monoclonal antibodies for western blotting, by MP Biomedicals (1 mentions)
C 18 ziptip columns, by Merck Group (1 mentions)
Ficoll histopaque density gradient, by Merck Group (1 mentions)
Dithiothreitol (dtt), by Bio-Rad (1 mentions)
Ipg strip, by Bio-Rad (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
3 protocols using polyvinylidene difluoride pvdf membrane 0.45 μm
1
Immunomodulatory Effects of Gymnema Sylvestre
2
Comprehensive Proteomics Sample Preparation
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All reagents used were of molecular grade, protease inhibitor cocktail, Phenylmethylsulfonyl fluoride (PMSF), Proteomics grade trypsin from porcine pancreas, Iodoacetic acid (IAA), Acrylamide, bis Acrylamide, ficoll histopaque density gradient and Ammonium bicarbonate were procured from Sigma-Aldrich, St. Louis, MO, USA. Sodium Dodecyl Sulfate (SDS), IPG strips, Urea, thioUrea, Mineral oil, Dithiothreitol (DTT), CHAPS (3-((3-cholamidopropyl) dimethylammonio)-1-propanesulfonate) were purchased from Bio-Rad Laboratories, Inc. California, USA. For protein quantification, Pierce™ 660nm Protein Assay kit from Thermo Fisher Scientific Inc. USA was used. Mouse, Anti-AGE monoclonal antibodies for western blotting were obtained from MP Biomedicals, USA. The polyvinylidenedifluoride (PVDF) membrane (0.45μm) and dialysis tubing were purchased from Millipore Corporation, MA, USA. C-18 ZipTip columns for concentrating and desalting the peptides were purchased from Millipore, Billerica, MA, USA. Pre-stained protein ladder was procured from Real Biotech Corporation, Taipei, Taiwan.
3
Immunoblotting Protocol for Protein Analysis
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For immunoblotting, cells were lysed in sample buffer (0.125 M Tris–HCl pH 6.8, 2% SDS, 0.005% bromophenol blue, 21% glycerol, 4% β-mercaptoethanol) and boiled for 5 min at 98°C. Lysates were separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) using NuPAGE™ 4–12% Bis–Tris gels (Invitrogen) and transferred to polyvinylidene difluoride (PVDF) membrane (0.45 μm) (Millipore). Membranes were blocked in 5% BSA (Sigma) for 30 min at room temperature and incubated for 2 h at room temperature or overnight at 4°C with primary antibodies against GFP (Roche), androgen receptor (Invitrogen), PTK2 (Invitrogen), XPB (Abcam), CSB (Bethyl), XPF (Santa Cruz), XPC (Bethyl), Tubulin (Sigma Aldrich) or Ku70 (Santa Cruz). Membranes were incubated with secondary antibodies coupled to IRDyes (Sigma) and scanned using an Odyssey CLx infrared scanner (LiCor). Antibodies, identifiers and concentrations are listed in Supplementary Table S4 .
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