To determine the partial amino acid sequences of sample proteins, in-gel digestion was performed according to the method of Cleveland et al. (1977) (link) using Staphylococcus aureus V8 protease (Fujifilm Wako Pure Chemical Industries, Ltd.) as described previously (Maehashi et al., 2007 (link)). After electroblotting onto a PVDF membrane (Bio-Rad Laboratories, Hercules, CA, USA), the separated band of a proteolytic fragment was excised from the membrane and subjected to amino (N)-terminal sequence (30 cycles) analysis using a gas-phase protein sequencer (PPSQ-21; Shimadzu, Kyoto, Japan). By using the first 20 amino acid sequences data were analyzed using NCBI’s protein BLAST program.
Staphylococcus aureus v8 protease
Manufactured by Fujifilm
Sourced in Japan
Staphylococcus aureus V8 protease is a laboratory enzyme used for protein analysis. It is a serine protease that cleaves peptide bonds on the C-terminal side of glutamic acid residues in protein sequences.
Automatically generated - may contain errors
Lab products found in correlation
Ppsq 21, by Shimadzu (1 mentions)
Pvdf membrane, by Bio-Rad (1 mentions)
Resource s column, by GE Healthcare (1 mentions)
Sephadex g 15, by GE Healthcare (1 mentions)
Sepharose 4b, by GE Healthcare (1 mentions)
Mucin from porcine stomach type 2, by Merck Group (1 mentions)
Tskgel ods 120t, by Tosoh (1 mentions)
2 protocols using staphylococcus aureus v8 protease
1
Protein Sequence Determination by V8 Protease Digestion
2
Isolation and Characterization of Pearl Shell Proteins
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Cultured large winged pearl shells (Pteria penguin, 6-years-old) and their larva were kindly provided by Amami South Sea & Mabe pearl Co. Ltd. (former Amami-branch, Tasaki & Co. Ltd.), Kagoshima, Japan. The mantle was collected and stored at −80°C until use. Resource S column, Sephadex G-15 and Sepharose 4B gels were purchased from GE Healthcare (NJ, USA). TSKgel sugar AXI, TSKgel ODS 120T, and TSKgel Amide 80 columns were from Tosoh (Tokyo, Japan). Trehalose-Sepharose 4B gel was prepared according to the method of Matsumoto et al. [29] . Mucin type I from bovine submaxillary glands and mucin type II from porcine stomach were purchased from Sigma Chemical (St. Louis, MO, USA). Achromobacter protease I and Staphylococcus aureus V8 protease were purchased from Wako Pure Chemical (Osaka, Japan). Glycopeptidase A was purchased from Seikagaku Kogyo (Tokyo, Japan). Trehalose was purchased from Hayashibara (Okayama, Japan). All the other reagents were of the purest grade commercially available.
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