A complex mixture of peptides produced by Trypsin digestion of Bovine Serum Albumin (BSA) that was reduced and alkylated with Iodoacetamide (CAM modified). This peptide mixture can be used to test a Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight (MALDI-TOF) or Electrospray Ionization (ESI) mass spectrometer (TOF, Q-TOF or Ion Trap).
MALDI Analysis of BSA Digest:
The BSA digest is dissolved to 1 pmol/μl. One µl of digest is mixed with 1 µl of α-cyano-4-hydroxycinnamic acid (10 mg/ml in 50:50 acetonitrile:water with 0.1% trifluoroacetic acid). One μl of the digest/matrix solution were then spotted directly onto a MALDI target plate. Online Analysis of BSA Digest: The BSA digest solution was diluted to 100 fmol/μl with 0.1% formic acid. One μl (100 fmol) of the digest solution was injected via an Easy-nanoLC II (Proxeon) onto a self-packed reverse phase C18 nano column (Phenomenex packing material, 5 cm, 150 μm) and loaded onto a self-packed C18 analytical column with an integrated tip (New Objective Picofrit, 15 cm, 100 μm, Phenomenex packing material). Peptides were separated using a 45 min 5-70%B gradient (A = 0.1% formic acid, B = CH3CN, 0.1% formic acid) at a flow rate of 400 nl/min. Eluting peptides were analyzed online by a Q-Exactive mass spectrometer (Thermo Scientific) with an electrospray ion source. MS/MS spectra were acquired by HCD using an acquisition range of 400 to 1600 m/z.
Product Source
BSA (GENBANK P02769) was digested using Trypsin (TPCK-treated).
Reconstitution
Suggested volume to resuspend: 500 µl. Avoid repeated freeze/thaw cycles once in solution.
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