Monoclonal Antibodies Raised against Carp Myosin

Nakaya, Misako; Watabe, Shugo

doi:10.2331/fishsci.60.47

Cited by 3 publications

(3 citation statements)

References 35 publications

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“…The cDNA library was screened using anti‐carp myosin polyclonal antibody 9 by the method of Sambrook et al 10 Vector pBluescript SK − and Escherichia coli strain XL1‐Blue were purchased from Stratagene. Escherichia coli infected with the λZAP II cDNA library was cultured on an agar plate at 42°C for 4 h, and a nitrocellulose filter containing 10 mM isopropyl‐β‐ D ‐thiogalactopyranoside was overlaid to the plate and incubated at 37°C for 4 h. The nitrocellulose filter replica was screened with anticarp myosin polyclonal antibody.…”

Section: Methodsmentioning

confidence: 99%

cDNA cloning of myosin heavy chain from white croaker fast skeletal muscle and characterization of its complete primary structure

et al. 2000

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SUMMARY: A cDNA library constructed from the dorsal fast skeletal muscle of white croaker Pennahia argentata was screened for myosin heavy chain using antibody raised against carp fast skeletal myosin. A full‐length cDNA was further cloned by reverse transcription–PCR and 5′‐RACE using first‐strand cDNA as a template together with appropriate sets of primers. The entire cDNA consisted of 5986 nucleotides (nt) with 64 nt 5′‐untranslated and 129 nt 3′‐untranslated regions. This full‐length cDNA had an open reading frame encoding a polypeptide of 1930 amino acid residues. Amino acid alignment with myosins from various vertebrates revealed some striking differences between fish and mammalian sequences which could be due to their position in the vertebrate evolutionary process. Hydrophilicity analysis revealed two different features of the myosin molecule: S1 heavy chain showed a mixed profile of hydrophobicity and hydrophilicity, whereas rod had a profile of only hydrophilicity. Comparison of amino acid sequences in the rod region between white croaker and walleye pollack showed a markedly high identity of 92%. White croaker myosin rod had a characteristic seven‐residue (heptad) repeat (a, b, c, d, e, f, g)n, where positions a and d were normally occupied by hydrophobic residues, and positions b, c and f by oppositely charged residues, which may lead to interhelical electrostatic attractions stabilizing the coiled‐coil of α‐helices.

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Section: Methodsmentioning

confidence: 99%

cDNA cloning of myosin heavy chain from white croaker fast skeletal muscle and characterization of its complete primary structure

et al. 2000

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“…This transferring time was not enough for blotting MHC, but it was enough for bands less than 170 kDa. After the blocking, the membrane was incubated in the 1st antibody diluted 1:40 in the blocking solution at 4 ЊC for 16 h. The preceding treatment was made according to a common procedure (Nakaya and Watabe 1994).…”

Section: Transfer Condition and Western Blottingmentioning

confidence: 99%

Heat‐Induced Changes of Myosin and Sarcoplasmic Proteins in Beef During Simmering

Tajima¹,

Ito²,

Arakawa³

et al. 2001

Journal of Food Science

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Changes of proteins in beef cubes, soup stock, and scum (a complex of coagulated proteins, fat, and minerals floating on soup stock) during simmering in sub-boiling water at 95 8C for 3 h were investigated by SDS-PAGE and Western blotting. Of the soluble proteins in soup stock heated for 1 h, 3 components having molecular weights between 47 and 36 kDa reacted with myosin antibody. Those bands disappeared from soup stock after heating for 3 h, however, they were observed in the scum. Bands of 70 and 58 kDa, which reacted with myosin antibody and were observed after 1 h of heating at 95 8C, were also observed after 30 min of heating under pressure.

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“…The reactions were run for 0.5 to 5 min and then terminated by adding phenylmethylsulfonyl fluoride (PMSF) to a final concentration of 1 mM. Myosin rod was digested with ␣-chymotrypsin according to the procedure of Nakaya and Watabe (1994) with a slight modification.…”

Section: Proteolytic Digestion Of Myosin S-1 and Rodmentioning

confidence: 99%

Monitoring Myosin Degradation During Conditioning in Chicken Meat Using an Immunological Method

Ikeuchi

Kamiyama

Suzuki

et al. 2001

Journal of Food Science

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Changes of chicken breast myosin during storage at 2 ЊC and 37 ЊC were monitored immunochemically. Anti-myosin subfragment-1 (S-1) monoclonal antibody, which recognized epitopes within the 27 kDa fragment of S-1, and the anti-myosin rod polyclonal antiserum, were prepared. Myosin degradation products were not detected in muscle extracts stored for 3 weeks at 2 ЊC. In contrast, storage at 37 ЊC brought about the degradation of myosin heavy chain to immunologically detectable small fragments. While, myosin rod produced during the conditioning period was not decomposed into any small filaments. Namely, storage of muscle at 37 ЊC resulted in minor amounts of myosin heavy chain degradation, with initial conversion to rod and S-1 fragments, and subsequent breakdown occurred in the S-1 region only. Immunoblot assay also suggested that the pattern of changes in myosin heavy chain in muscle incubated at 37 ЊC was similar to that produced by in vitro digestion with cathepsin D.

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Monoclonal Antibodies Raised against Carp Myosin

Cited by 3 publications

References 35 publications

cDNA cloning of myosin heavy chain from white croaker fast skeletal muscle and characterization of its complete primary structure

cDNA cloning of myosin heavy chain from white croaker fast skeletal muscle and characterization of its complete primary structure

Heat‐Induced Changes of Myosin and Sarcoplasmic Proteins in Beef During Simmering

Monitoring Myosin Degradation During Conditioning in Chicken Meat Using an Immunological Method

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