Ammonium bicarbonate, dithiothreitol (DTT), and iodoacetamide (IAA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). MS grade trypsin protease, Spectra™ Multicolor Broad Range Protein Ladder (10 to 260 kDa, catalog number: 26634), and HPLC grade solvents were purchased from Thermo Scientific™ Pierce™ (Rockford, IL, USA). Acetonitrile (HPLC grade), LiChrospher® WP 300 C18 RP-HPLC column, and Millipore ZipTip® C18 Pipette Tips were obtained from Merck (Kenilworth, NJ, USA). Other chemicals and solvents used were of analytical grade and purchased from Sigma-Aldrich (USA).
Millipore ziptip c18 pipette tips
Manufactured by Merck Group
Sourced in United States
The Millipore ZipTip® C18 Pipette Tips are a type of pipette tip designed for sample preparation in analytical chemistry and biochemistry applications. They feature a small C18 stationary phase material packed within the tip, which allows for the extraction and purification of analytes from complex samples prior to instrumental analysis.
Automatically generated - may contain errors
Lab products found in correlation
Ammonium bicarbonate, by Merck Group (10 mentions)
Iodoacetamide, by Merck Group (9 mentions)
Dithiothreitol (dtt), by Merck Group (8 mentions)
Spectra multicolor broad range protein ladder, by Thermo Fisher Scientific (3 mentions)
Ms grade trypsin protease, by Thermo Fisher Scientific (3 mentions)
Hplc grade solvents, by Thermo Fisher Scientific (3 mentions)
Spectra multicolor broad range protein ladder 10 to 260 kda, by Thermo Fisher Scientific (2 mentions)
Trifluoroacetic acid, by Thermo Fisher Scientific (1 mentions)
Hplc grade acetonitrile acn, by Thermo Fisher Scientific (1 mentions)
Lichrospher wp 300 rp 18, by Merck Group (1 mentions)
Pageruler prestained protein ladder, by Thermo Fisher Scientific (1 mentions)
13 protocols using millipore ziptip c18 pipette tips
1
Proteomic Sample Preparation Protocol
2
Proteomics Sample Preparation Protocol
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Ammonium bicarbonate, dithiothreitol (DTT) and iodoacetamide were purchased from Sigma-Aldrich (USA). Mass-spectrometry grade trypsin protease, Spectra™ Multicolor Broad Range Protein Ladder (10 to 260 kDa), and HPLC grade solvents used in the studies were purchased from Thermo Scientific™ Pierce™ (USA). Millipore ZipTip® C18 Pipette Tips were obtained from Merck, USA. Other chemicals and reagents of analytical grade were purchased from Sigma-Aldrich (USA).
3
Venom Neutralization by Sea Snake Antivenom
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Hydrophis curtus venom was a pooled sample from 10 adult snakes collected in the waters of Penang Island (Malaysia). The antivenom tested was Sea Snake Antivenom (SSAV) produced by CSL Ltd. (Melbourne, Australia, Seqirus Ltd. currently). The antivenom was in (liquid form) contained F(ab’)2 from horses hyperimmunized against the venoms of Hydrophis schistosus (beaked sea snake, obtained from the waters of Penang Island, Malaysia) and the Australian Notechis scutatus (common tiger snake). The protein concentration of the antivenom was 217.2 ± 3.0 mg/mL, determined in a previous study from the laboratory [25 (link)].
The chemicals and reagents used were primarily supplied by Sigma-Aldrich (Saint Louis, MO, USA) and were of analytical grade. The molecular weight marker Spectra™ Multicolor Broad Range Protein Ladder (10–260 kDa), trypsin (mass spectrometry grade), HPLC grade acetonitrile (ACN) and trifluoroacetic acid (TFA) were purchased from Thermo Scientific™ Pierce™ (Waltham, MA, USA). Millipore ZipTip® C18 Pipette Tips and LiChrospher® WP 300 RP-18 (5 µm) were supplied by Merck (Burlington, MA, USA).
The chemicals and reagents used were primarily supplied by Sigma-Aldrich (Saint Louis, MO, USA) and were of analytical grade. The molecular weight marker Spectra™ Multicolor Broad Range Protein Ladder (10–260 kDa), trypsin (mass spectrometry grade), HPLC grade acetonitrile (ACN) and trifluoroacetic acid (TFA) were purchased from Thermo Scientific™ Pierce™ (Waltham, MA, USA). Millipore ZipTip® C18 Pipette Tips and LiChrospher® WP 300 RP-18 (5 µm) were supplied by Merck (Burlington, MA, USA).
4
Analytical Grade Reagents for Proteomic Analysis
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All chemicals and reagents used in this study were of analytical grade. Ammonium
bicarbonate, dithiothreitol (DTT) and iodoacetamide were purchased from
Sigma-Aldrich (USA). Mass spectrometry sequencing grade of trypsin proteases,
Thermo Scientific Spectra Multicolor Broad Range Protein Ladder (10-260 kDa) and
high-performance liquid chromatography (HPLC) grade solvents used in the studies
were purchased from Thermo Scientific™ Pierce™ (USA). Millipore ZipTip® C18
Pipette Tips were purchased from Merck (USA).
bicarbonate, dithiothreitol (DTT) and iodoacetamide were purchased from
Sigma-Aldrich (USA). Mass spectrometry sequencing grade of trypsin proteases,
Thermo Scientific Spectra Multicolor Broad Range Protein Ladder (10-260 kDa) and
high-performance liquid chromatography (HPLC) grade solvents used in the studies
were purchased from Thermo Scientific™ Pierce™ (USA). Millipore ZipTip® C18
Pipette Tips were purchased from Merck (USA).
5
Mass Spectrometry Proteomics Protocol
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All chemicals and reagents used in the studies were of analytical grade. Ammonium bicarbonate, dithiothreitol (DTT) and iodoacetamide were purchased from Sigma-Aldrich (USA). Mass spectrometry sequencing grade of trypsin and chymotrypsin proteases, Spectra™ Multicolor Low Range Protein Ladder (1.7 to 40 kDa), and HPLC grade solvents used in the studies were purchased from Thermo Scientific™ Pierce™ (USA). LiChroCART® 250-4 LiChrospher® WP 300 RP-18 (5 μm) HPLC cartridge and Millipore ZipTip® C18 Pipette Tips were purchased from Merck (USA).
6
Proteomic Analysis of Biological Samples
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All chemicals and reagents used were of analytical grade. Ammonium bicarbonate, dithiothreitol (DTT) and iodoacetamide were purchased from Sigma-Aldrich (St. Louis, MO, USA). MS grade trypsin protease, Spectra™ Multicolor Broad Range Protein Ladder (10 to 170 kDa), and HPLC grade solvents used in the studies were purchased from Thermo® Scientific Pierce (Waltham, MA, USA). Resource® S cation-exchange column (1 mL) was purchased from GE Healthcare (Uppsala, Sweden). LiChroCART® 250-4 LiChrospher® WP 300 and Millipore ZipTip® C18 Pipette Tips were purchased from Merck (Kenilworth, NJ, USA).
7
Protein Fractionation and Trypsin Digestion
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Fractionated proteins collected from the reverse-phase were reduced by DTT (Dithiothreitol), alkylated with IAA (iodoacetamide) and subjected to in-solution proteolytic digestion with trypsin according to the method reported previously [47 (link)]. Millipore ZipTip® C18 Pipette Tips (Merck, Burlington, MA, USA) were used to clean up and desalt the trypsin-digested peptides for an enhanced performance of mass spectrometry analysis.
8
SDS-PAGE and Mass Spectrometry Analysis of Purified Toxins
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The purified toxins were subjected to 15% sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) under reducing condition as described by Laemmli [34 (link)] and calibrated with the Thermo™ Scientific™ PageRuler Prestained Protein Ladder (10–170 kDa). Separately, the purified toxins were subjected to reduction with DTT, alkylation with iodoacetamide, and in-solution digestion with mass-spectrometry grade trypsin protease according to the manufacturer’s protocol (Thermo Scientific™ Pierce™). The trypsin-digested peptides were desalted with Millipore ZipTip® C18 Pipette Tips (Merck) according to the manufacturer’s protocol to enhance the performance of mass spectrometry.
9
Proteome Preparation and Mass Spectrometry
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All chemicals and materials used were of analytical grade. Ammonium bicarbonate, dithiothreitol (DTT) and iodoacetamide (IAA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). MS grade trypsin protease, Spectra™ Multicolor Broad Range Protein Ladder (Catalog number: 26634, 10 to 260 kDa), and HPLC grade solvents were purchased from Thermo Scientific™ (Pierce™, Rockford, IL, USA). Millipore ZipTip® C18 Pipette Tips were obtained from Merck KGaA (Darmstadt, Germany). Other chemicals and reagents of analytical grade were purchased from Sigma-Aldrich (St. Louis, MO, USA).
10
Protein Identification Protocol
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All chemicals and reagents used were of analytical grade. Ammonium bicarbonate, dithiothreitol (DTT), and iodoacetamide were purchased from Sigma-Aldrich (St. Louis, MO, USA). MS-grade trypsin protease was purchased from ThermoFisher Scientific (Waltham, MA, USA). Millipore ZipTip C18 pipette tips were purchased from Merck (Kenilworth, NJ, USA). PM2700 ExcelBand 3-color Broad Range Protein Marker (~5–245 kDa) was obtained from SMOBIO (Hsinchu, Taiwan).
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