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Table 5 Materials for QTAP

From: A study protocol for quantitative targeted absolute proteomics (QTAP) by LC-MS/MS: application for inter-strain differences in protein expression levels of transporters, receptors, claudin-5, and marker proteins at the blood–brain barrier in ddY, FVB, and C57BL/6J mice

Name Company Catalog# Amount Note
Guanidine HCl Wako 070-01825 500 g For denaturing buffer
2NA (EDTA·2Na) DOJINDO 345-01865 500 g For denaturing buffer
Trizma® (Tris) base, primary standard and buffer SIGMA T1503-1KG 1 kg For several buffers
Ammonium hydrogencarbonate Wako 017-02875 500 g For LysC and ProteaseMax solution
(±)-Dithiothreitol (DTT) Wako 049-08972 25 g  
Iodoacetamide (IAA) Wako 093-02152 25 g  
Chloroform Wako 038-02606 500 mL  
Methanol Wako 137-01823 3 L  
Urea Wako 211-01213 1 kg  
ProteaseMax surfactant, trypsin enhancer Promega V2072 5 × 1 mg  
Lysyl endopeptidase (LysC) Wako 129-02541 10 AU  
Sequencing grade modified trypsin, frozen (TPCK-trypsin) Promega V5113 100 μg  
SUMILON proteosave SS, 1.5 mL tube SUMITOMO BAKELITE MS-4215 M 100/pk Low protein binding tube
SUMILON proteosave SS, 0.5 mL tube SUMITOMO BAKELITE MS-4205 M 100/pk Low protein binding tube
Ultra plus tip 1–10 μL bms UP-0110 96/pk Low protein binding tip
Ultra plus tip 1–200 μL bms UP-2010 96/pk Low protein binding tip
Eyela cute mixer CM-1000 TOKYO RIKAKIKAI 188140   Tube mixer
Bransonic ultrasonic cleaner 2510 J-DTH Branson 2510 J-DTH   Sonicator
High speed refrigerated micro centrifuge MX-160 TOMY    Centrifuge
TMA-30 TOMY    Angle rotor
TMS-21 TOMY    Swing rotor
Block incubator BI-525 ASTEC    LysC digestion
Incubator MIR-262 SANYO MIR-262   Trypsin digestion
Formic acid (98.0%) Wako 066-00466 500 mL Acidification
Acetonitrile (LC/MS grade) Wako 018-19853 3 L Mobile phase
XBridge BEH130 C18 (1.0 × 100 mm, 3.5 μm) column Waters 186003561   HPLC column
XBridge C18 Guard Cartridge (2.1 × 10 mm, 3.5 μm) Waters 186003059 2/pk Guard column
Sentry 2.1 mm guard holder Waters WAT097958   Guard column holder
Agilent technologies 1200 series (HPLC system) Agilent    HPLC
API5000 AB Sciex    QqQ mass spectrometer
QTRAP5500 AB Sciex    QqQ mass spectrometer
96 well plates, 0.5 mL, polypropylene Agilent 5042-1386 10/pk Sample plate for auto sampler (HPLC)
Pre-slit well cap for 96 well PP plate non sterile, silicone Thermo SCIENTIFIC 276011 10/pk Sample plate cap for auto sampler (HPLC)
DC protein assay reagent A BIO-RAD 500-0113 250 mL Protein quantification (Lowry method)
DC protein assay reagent B BIO-RAD 500-0114 1 L Protein quantification (Lowry method)