1983
DOI: 10.1073/pnas.80.6.1506
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Peptide antibody specific for the amino terminus of skeletal muscle alpha-actin.

Abstract: The NH2-terminal peptide of skeletal muscle aactin (SaN peptide), which contains a primary sequence unique to this actin isozyme, was used to prepare an isozyme-specific peptide antibody. SaN peptide was purified from chicken breast muscle actin by preparative reverse-phase HPLC and was coupled to hemocyanin. This complex was used to immunize rabbits in order to elicit actin antibodies specific for the skeletal muscle a-actin isozyme. (1)(2)(3)(4), and these isozymes may account for some of actin's tissue-spe… Show more

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Cited by 71 publications
(57 citation statements)
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“…It has been shown the 3-and 7-actins and not a-actin are predominant in cultured skeletal msucle cells before fusion and that a-actin appears after fusion (5,20,45). Moreover, an antibody that recognizes cytoplasmic actin (probably 3-and 3,-actins) but not myofibriUar actin (a-actin) stains the cytoplasm of fully differentiated skeletal muscle fibers diffusely and the membrane-associated structures strongly (32).…”
Section: Discussionmentioning
confidence: 99%
“…It has been shown the 3-and 7-actins and not a-actin are predominant in cultured skeletal msucle cells before fusion and that a-actin appears after fusion (5,20,45). Moreover, an antibody that recognizes cytoplasmic actin (probably 3-and 3,-actins) but not myofibriUar actin (a-actin) stains the cytoplasm of fully differentiated skeletal muscle fibers diffusely and the membrane-associated structures strongly (32).…”
Section: Discussionmentioning
confidence: 99%
“…The protein was eluted by 0.15 M NaCl, 50 mM sodium phosphate pH 7.0. The absorbance of the fractions at 280nm was measured and those containing the modified ovalbumin were mixed with 10 mg peptide 567-574 for 3 h. Peptide 238-246 was coupled to Limulus hemocyanin essentially by the glutaraldehyde method of Bulinski et al (1983). The coupled peptides were separated by SDSPAGE to verify the coupling.…”
Section: Methodsmentioning
confidence: 99%
“…After 3 months, one rabbit exhibited a low titer and was boosted every 2 to 3 months with 3 mg of CK prepared as follows. One-third of the CK was native, onethird was denatured with 1% sodium dodecyl sulfate (SDS) and embedded in a 10%o polyacrylamide gel, and one-third was cross-linked with glutaraldehyde as described by Bulinski et al (8). After homogenization, this mixture was emulsified with an equal volume of Freund complete adjuvant and injected subcutaneously along the back and neck.…”
Section: Methodsmentioning
confidence: 99%
“…Antibody titer, cross-reactivity with other CK isozymes, and specificity of binding was demonstrated by using an immunoblot technique (61) modified as described before (8). Tissue extracts were made for the immunoblot assay by homogenizing adult skeletal muscle or brain directly into SDS gel sample buffer (prepared as described by Laemmli [34] myoblasts were grown and induced to differentiate as previously described (36) except that Ham's F-10 medium containing 1.1 mM CaCl2 (F-10C), 1 ,uM insulin, 100 nM dexamethasone, and 5% horse serum was used in place of conditioned medium.…”
Section: Methodsmentioning
confidence: 99%